Macrocyclic compounds useful as pharmaceuticals

ABSTRACT

The present invention provides compositions comprising compounds having formula (I):  
                 
 
     and additionally provides methods for the use thereof in the treatment of various disorders including inflammatory or autoimmune disorders, and disorders involving malignancy or increased angiogenesis, wherein R 1 -R 11 , X, Y, Z, and n are as defined herein. In certain embodiments, the compositions are for systemic (e.g., oral) administration. In certain embodiments, methods for the treatment of various disorders including inflammatory or autoimmune disorders comprise systemically (e.g., orally) administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I).

PRIORITY CLAIM

[0001] This Application claims the benefit under 35 U.S.C. § 120 ofInternational Application No.: PCT/US03/07377, filed Mar. 7, 2003, whichclaims priority to U.S. Provisional Patent Application Nos. 60/362,883,filed Mar. 8, 2002, and 60/380,711, filed May 14, 2002; each of theseapplications is hereby incorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION

[0002] F152 (LL-Z1640-2) (1) is a zearalenone-like macrolide, isolatedfrom shake flask fermentation, crude extracts of which inhibited theciliated protozoan Tetrahymena pyriformis (see, McGahren et al. J. Org.Chem. 1978, 43, 2339). It was reported that initial biological studiesusing this natural product failed to yield any particularly interestingactivities.

[0003] After initial isolation and reporting of this compound, severalother groups explored the possibility of preparing additionalderivatives and/or further exploring their biological activity. Forexample, scientists at Merck reported that F152 and certain isomersthereof inhibit the phosphorylating enzyme Map/Erk kinase (MEK) and thusare useful for the treatment of certain cancers and other diseasescharacterized by the formation of neoangiogenesis (see, GB 323 845).Other groups have also reported derivatives of F152 having activity astyrosine kinase inhibitors, which are useful, for example, for thetreatment of cancer and inflammatory disorders (see, EP 606 044; WO00/38674; JP 8-40893; WO 96/13259; U.S. Pat. Nos. 5,728,726; 5,674,892;5,795,910). Each of these groups, however, was only able to obtain F152and derivatives thereof by fermentation techniques and by modificationsto the natural product, respectively, and thus were limited in thenumber and types of derivatives that could be prepared and evaluated forbiological activity. Additionally, although F152 and certain derivativesthereof have demonstrated potent in vitro activities, these compoundsare biologically unstable (for example, they are susceptible to enoneisomerization in mouse and human plasma), thereby limiting thedevelopment of these compounds as therapeutics for the treatment ofhumans or other animals.

[0004] Clearly, there remains a need to develop synthetic methodologiesto access and examine the therapeutic effect of a variety of novelanalogues of F152, particularly those that are inaccessible by makingmodifications to the natural product. It would also be of particularinterest to develop novel compounds that exhibit a favorable therapeuticprofile in vivo (e.g., are safe and effective, while retaining stabilityin biological media).

SUMMARY OF THE INVENTION

[0005] As discussed above, there remains a need for the development ofnovel analogues of F152 and the evaluation of their biological activity.The present invention provides compositions for the systemicadministration of compounds of general formula (I):

[0006] and a pharmaceutically acceptable carrier or diluent, whereinR₁-R₁₁, X, Y, Z, and n are as defined generally and in classes andsubclasses herein. In certain embodiments, the compositions are for oraladministration. In certain other embodiments, the compound is present inan amount effective to inhibit production of a pro-inflammatory and/orimmunologic cytokine. In certain exemplary embodiments, the citokine isTNFα, IL-1, IL-6, IL-8 or IL-2.

[0007] In another aspect, the invention provides a method for the use ofcompounds of formula (I) in the treatment of various disorders includinginflammatory or autoimmune disorders via systemic administration ofthese compounds. In certain embodiments, the method comprisessystemically (e.g., orally) administering to a subject in need thereof atherapeutically effective amount of a compound of formula (I). Incertain other embodiments, the compound is present in an amounteffective to inhibit production of a pro-inflammatory and/or immunologiccytokine. In certain exemplary embodiments, the citokine is TNFα, IL-1,IL-6, IL-8 or IL-2. In certain other embodiments, the method is fortreating psoriasis.

DESCRIPTION OF CERTAIN PREFERRED EMBODIMENTS OF THE INVENTION

[0008] In recognition of the need to access and further explore thebiological activity of novel analogues of F152, and this class ofmacrocycles in general, the present invention provides novel macrocycliccompounds, as described in more detail herein, which demonstrateincreased stability and are potent inhibitors of NF-κB activation, AP-1activation and protein kinases (for example, MEKK, MEK1, PDGFr, VEGFr).Based on these mechanisms of action, the compounds inhibit theproduction of various pro-inflammatory and/or immunologic cytokines suchas TNFα, IL-1, IL-6, IL-8, IL-2 etc, and also inhibit the production ofvarious pro-inflammatory molecules under the regulation of NF-κB pathwaysuch as prostaglandins produced from COX-2, ICAM-1 and MMP-1 and 3 etc.Also, the compounds have ability to inhibit cell proliferation under theregulation of AP-1 pathway through the inhibition of MEK1. In addition,the compounds have ability to inhibit angiogenesis mainly based on theinhibitory activities on VEGFr and PDGFr kinases. Thus, the compounds ofthe invention, and pharmaceutical compositions thereof, are useful asanti-inflammatory and/or immunosuppressive agents for the treatment ofvarious inflammatory diseases, and abnormal cell proliferation or asantiangiogenesis agents for the treatment of cancer. In certainembodiments, the compounds of the present invention can be used for thetreatment of diseases and disorders including, but not limited tosepsis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis,inflammatory bowel disease (Crohn's disease and ulcerative colitis),multiple sclerosis, atopic dermatitis, psoriasis, asthma, osteoporosis,allergic rhinitis, ocular inflammation, hepatitis, autoimmune disorders,systemic lupus erthematosus, allograft rejection/graft versus hostdisease, diabetes, AIDS, solid tumor cancers, leukemia, lymphomas,non-hodgkin's B-cell lymphomas, chronical lymphocytic leukemia (CLL),multiple myeloma, eczema, urticaria, myasthenia gravis, idiopathicthrombocytopenia purpura, cardiovascular disease (e.g., myocardialinfarction, atherosclerosis), hepatitis, glomerulonephropathy,productive nephritis, adenovirus, diseases/disorders of the centralnervous system (e.g., stroke, Alzheimer's disease, epilepsy) and for thetreatment of the symptoms of malaria, to name a few. The inventivecompounds also find use in the prevention of restenosis of blood vesselssubject to traumas such as angioplasty and stenting.

[0009] In addition, it has been shown that photoaging of undamaged skindue to UVB irradiation exposure is inhibited by administering an agentthat inhibits one or both of the transcription factors AP-1 and NF-κB tothe skin prior to such exposure (See, for example, U.S. Pat. No.5,837,224). Therefore, the inventive compounds, and pharmaceuticalcompositions thereof, are useful in the treatment of photoaging-relateddisorders/conditions.

[0010] 1) General Description of Compounds of the Invention

[0011] In certain embodiments, the compounds of the invention includecompounds of the general formula (I) as further defined below:

[0012] wherein R₁ is hydrogen, aliphatic, heteroaliphatic, alicyclic,heteroalicyclic, aryl or heteroaryl;

[0013] R₂ and R₃ are each independently hydrogen, halogen, hydroxyl,protected hydroxyl, or an aliphatic, heteroaliphatic, alicyclic,heteroalicyclic, aryl or heteroaryl moiety; or

[0014] R₁ and R₂, when taken together, may form a substituted orunsubstituted, saturated or unsaturated cyclic ring of 3 to 8 carbonatoms; or

[0015] R₁ and R₃, when taken together, may form a substituted orunsubstituted, saturated or unsaturated cyclic ring of 3 to 8 carbonatoms;

[0016] R₄ is hydrogen or halogen;

[0017] R₅ is hydrogen, an oxygen protecting group or a prodrug;

[0018] R₆ is hydrogen, hydroxyl, or protected hydroxyl;

[0019] n is 0-2;

[0020] R₇, for each occurrence, is independently hydrogen, hydroxyl, orprotected hydroxyl;

[0021] R₈ is hydrogen, halogen, hydroxyl, protected hydroxyl, alkyloxy,or an aliphatic moiety optionally substituted with hydroxyl, protectedhydroxyl, SR₁₂, or NR₁₂R₁₃;

[0022] R₉ is hydrogen, halogen, hydroxyl, protected hydroxyl, OR₁₂,SR₁₂, NR₁₂R₁₃, —X₁(CH₂)_(p)X₂—R₁₄, or is lower alkyl optionallysubstituted with hydroxyl, protected hydroxyl, halogen, amino, protectedamino, or —X₁(CH₂)_(p)X₂—R₁₄;

[0023] wherein R₁₂ and R₁₃ are, independently for each occurrence,hydrogen, aliphatic, heteroaliphatic, alicyclic, heteroalicyclic, arylor heteroaryl; or a protecting group, or R₁₂ and R₁₃, taken together mayform a saturated or unsaturated cyclic ring containing 1 to 4 carbonatoms and 1 to 3 nitrogen or oxygen atoms, and each of R₁₂ and R₁₃ areoptionally further substituted with one or more occurrences of hydroxyl,protected hydroxyl, alkyloxy, amino, protected amino, alkylamino,aminoalkyl, or halogen,

[0024] wherein X₁ and X₂ are each independently absent, or are oxygen,NH, or —N(alkyl), or wherein X₂—R₁₄ together are N₃ or are a saturatedor unsaturated heterocyclic moiety,

[0025] p is 2-10, and

[0026] R₁₄ is hydrogen, or an aryl, heteroaryl, alkylaryl, oralkylheteroaryl moiety, or is —(C═O)NHR₁₅—(C═O)OR₁₅, or —(C═O)R₁₅,wherein each occurrence of R₁₅ is independently hydrogen, aliphatic,heteroaliphatic, alicyclic, heteroalicyclic, aryl or heteroaryl; or R₁₄is —SO₂(R₁₆), wherein R₁₆ is an aliphatic moiety, wherein one or more ofR₁₄, R₁₅, or R₁₆ are optionally substituted with one or more occurrencesof hydroxyl, protected hydroxyl, alkyloxy, amino, protected amino,alkylamino, aminoalkyl, or halogen; or

[0027] R₈ and R₉ may, when taken together, form a saturated orunsaturated cyclic ring containing 1 to 4 carbon atoms and 1 to 3nitrogen or oxygen atoms and is optionally substituted with hydroxyl,protected hydroxyl, alkyloxy, amino, protected amino, alkylamino,aminoalkyl, or halogen;

[0028] R₁₀ is hydrogen, hydroxyl, protected hydroxyl, amino, orprotected amino;

[0029] R₁₁ is hydrogen, hydroxyl or protected hydroxyl;

[0030] X is absent or is O, NH, N-alkyl, CH₂ or S;

[0031] Y is CHR₁₇, O, C═O, CR₁₇ or NR₁₇; and Z is CHR₁₈, O, C═O, CR₁₈ orNR₁₈, wherein each occurrence of R₁₇ and R₁₈ is independently hydrogenor aliphatic, or R₁₇ and R₁₈ taken together is —O—, —CH₂— or —NR₁₉—,wherein R₁₉ is hydrogen or lower alkyl, and Y and Z may be connected bya single or double bond; and

[0032] pharmaceutically acceptable derivatives thereof.

[0033] In certain embodiments of compounds described directly above andcompounds as described in certain classes and subclasses herein, thefollowing groups do not occur simultaneously as defined:

[0034] X is oxygen,

[0035] R₁ is methyl with S-configuration,

[0036] R₂ and R₃ are each hydrogen,

[0037] R₄ is hydrogen,

[0038] R₅ is hydrogen, lower alkyl or lower alkanoyl,

[0039] R₆ is OR′, where R′ is hydrogen, lower alkyl or lower alkanoylwith S-configuration,

[0040] R₇ is hydrogen,

[0041] Y and Z together represent —CHR₁₇—CHR₁₈— or —CR₁₇═CR₁₈—, whereinR₁₇ and R₁₈ are independently hydrogen, or when Y and Z are—CHR₁₇—CHR₁₈, R₁₇ and R₁₈ taken together are —O—;

[0042] R₈ is hydrogen or OR′, where R′ is hydrogen, lower alkyl or loweralkanoyl,

[0043] R₉ is OR′, where R′ is hydrogen, lower alkyl or lower alkanoyl,

[0044] R₁₀ is OR″, where R″ is hydrogen, lower alkyl or lower alkanoyl;and

[0045] R¹¹ is hydrogen.

[0046] In certain other embodiments, compounds of formula (I) aredefined as follows:

[0047] R₁ is hydrogen, straight or branched lower alkyl, straight orbranched lower heteroalkyl, or aryl,

[0048] wherein the alkyl, heteroalkyl, and aryl groups may optionally besubstituted with one or more occurrences of halogen, hydroxyl orprotected hydroxyl;

[0049] R₂ and R₃ are each independently hydrogen, halogen, hydroxyl,protected hydroxyl, straight or branched lower alkyl, straight orbranched lower heteroalkyl, or aryl,

[0050] wherein the alkyl, heteroalkyl, and aryl groups may optionally besubstituted with one or more occurrences of halogen, hydroxyl orprotected hydroxyl; or

[0051] R₁ and R₂, when taken together, may form a saturated orunsaturated cyclic ring of 3 to 8 carbon atoms, optionally substitutedwith one or more occurrences of halogen; or

[0052] R₁ and R₃, when taken together, may form a saturated orunsaturated cyclic ring of 3 to 8 carbon atoms, optionally substitutedwith one or more occurrences of halogen;

[0053] R₄ is hydrogen or halogen;

[0054] R₅ is hydrogen or a protecting group;

[0055] R₆ is hydrogen, hydroxyl, or protected hydroxyl;

[0056] n is 0-2;

[0057] R₇, for each occurrence, is independently hydrogen, hydroxyl, orprotected hydroxyl;

[0058] R₈ is hydrogen, halogen, hydroxyl, protected hydroxyl, alkyloxy,or lower alkyl optionally substituted with hydroxyl, protected hydroxyl,SR₁₂, or NR₁₂R₁₃;

[0059] R₉ is hydrogen, halogen, hydroxyl, protected hydroxyl, OR₁₂,SR₁₂, NR₁₂R₁₃, —X₁(CH₂)_(p)X₂—R₁₄, or is lower alkyl optionallysubstituted with hydroxyl, protected hydroxyl, halogen, amino, protectedamino, or —X₁(CH₂)_(p)X₂—R₁₄;

[0060] wherein R₁₂ and R₁₃ are, independently for each occurrence,hydrogen, lower alkyl, aryl, heteroaryl, alkylaryl, or alkylheteroaryl,or a protecting group, or R₁₂ and R₁₃, taken together may form asaturated or unsaturated cyclic ring containing 1 to 4 carbon atoms and1 to 3 nitrogen or oxygen atoms, and each of R₁₂ and R₁₃ are optionallyfurther substituted with one or more occurrences of hydroxyl, protectedhydroxyl, alkyloxy, amino, protected amino, alkylamino, aminoalkyl, orhalogen,

[0061] wherein X₁ and X₂ are each independently absent, or are oxygen,NH, or —N(alkyl), or wherein X₂—R₁₄ together are N₃ or are a saturatedor unsaturated heterocyclic moiety,

[0062] p is 2-10, and

[0063] R₁₄ is hydrogen, or an aryl, heteroaryl, alkylaryl, oralkylheteroaryl moiety, or is —(C═O)NHR₁₅—(C═O)OR₁₅, or —(C═O)R₁₅,wherein each occurrence of R₁₅ is independently hydrogen, alkyl,heteroalkyl, aryl, heteroaryl, alkylaryl, or alkylheteroaryl, or R₁₄ is—SO₂(R₁₆), wherein R₁₆ is an alkyl moiety, wherein one or more of R₁₄,R₁₅, or R₁₆ are optionally substituted with one or more occurrences ofhydroxyl, protected hydroxyl, alkyloxy, amino, protected amino,alkylamino, aminoalkyl, or halogen; or

[0064] R₈ and R₉ may, when taken together, form a saturated orunsaturated cyclic ring containing 1 to 4 carbon atoms and 1 to 3nitrogen or oxygen atoms and is optionally substituted with hydroxyl,protected hydroxyl, alkyloxy, amino, protected amino, alkylamino,aminoalkyl, or halogen;

[0065] R₁₀ is hydrogen, hydroxyl, protected hydroxyl, amino, orprotected amino;

[0066] R₁₁ is hydrogen, hydroxyl or protected hydroxyl;

[0067] X is absent or is O, NH, N-alkyl, CH₂ or S;

[0068] Y is CHR₁₇, O, C═O, CR₁₇ or NR₁₇; and Z is CHR₁₈, O, C═O, CR₁₈ orNR₁₈, wherein each occurrence of R₁₇ and R₁₈ is independently hydrogenor lower alkyl, or R₁₇ and R₁₈ taken together is —O—, —CH₂— or —NR₁₉—,wherein R₁₉ is hydrogen or lower alkyl, and Y and Z may be connected bya single or double bond; and

[0069] pharmaceutically acceptable derivatives thereof.

[0070] In certain other embodiments, compounds of formula (I) aredefined as follows:

[0071] R₁ is hydrogen, straight or branched lower alkyl, straight orbranched lower heteroalkyl, or aryl,

[0072] wherein the alkyl, heteroalkyl, and aryl groups may optionally besubstituted with one or more occurrences of halogen, hydroxyl orprotected hydroxyl;

[0073] R₂ and R₃ are each independently hydrogen, halogen, hydroxyl,protected hydroxyl, straight or branched lower alkyl, straight orbranched lower heteroalkyl, or aryl,

[0074] wherein the alkyl, heteroalkyl, and aryl groups may optionally besubstituted with one or more occurrences of halogen, hydroxyl orprotected hydroxyl; or

[0075] R₁ and R₂, when taken together, may form a saturated orunsaturated cyclic ring of 3 to 8 carbon atoms, optionally substitutedwith one or more occurrences of halogen;

[0076] R₄ is hydrogen or halogen;

[0077] R₅ is hydrogen or a protecting group;

[0078] R₆ is hydrogen, hydroxyl, or protected hydroxyl;

[0079] n is 0-2;

[0080] R₇, for each occurrence, is independently hydrogen, hydroxyl, orprotected hydroxyl;

[0081] R₈ is hydrogen, halogen, hydroxyl, protected hydroxyl, alkyloxy,or lower alkyl optionally substituted with hydroxyl or protectedhydroxyl;

[0082] R₉ is hydrogen, halogen, hydroxyl, protected hydroxyl, OR₁₂,NR₁₂R₁₃, —X₁(CH₂)_(p)X₂—R₁₄, or is lower alkyl optionally substitutedwith hydroxyl, protected hydroxyl, halogen, amino, protected amino, or—X₁(CH₂)_(p)X₂—R₁₄;

[0083] wherein R₁₂ and R₁₃ are, independently for each occurrence,hydrogen, lower alkyl, aryl, heteroaryl, alkylaryl, or alkylheteroaryl,or a protecting group, or R₁₂ and R₁₃, taken together may form asaturated or unsaturated cyclic ring containing 1 to 4 carbon atoms and1 to 3 nitrogen or oxygen atoms, and each of R₁₂ and R₁₃ are optionallyfurther substituted with one or more occurrences of hydroxyl, protectedhydroxyl, alkyloxy, amino, protected amino, alkylamino, aminoalkyl, orhalogen, wherein X₁ and X₂ are each independently absent, or are oxygen,NH, or —N(alkyl), or wherein X₂—R₁₄ together are N₃ or are a saturatedor unsaturated heterocyclic moiety,

[0084] p is 2-10, and

[0085] R₁₄ is hydrogen, or an aryl, heteroaryl, alkylaryl, oralkylheteroaryl moiety, or is —(C═O)NHR₁₅—(C═O)OR₁₅, or —(C═O)R₁₅,wherein each occurrence of R₁₅ is independently hydrogen, alkyl,heteroalkyl, aryl, heteroaryl, alkylaryl, or alkylheteroaryl, or R₁₄ is—SO₂(R₁₆), wherein R₁₆ is an alkyl moiety, wherein one or more of R₁₄,R₁₅, or R₁₆ are optionally substituted with one or more occurrences ofhydroxyl, protected hydroxyl, alkyloxy, amino, protected amino,alkylamino, aminoalkyl, or halogen; or

[0086] R₈ and R₉ may, when taken together, form a saturated orunsaturated cyclic ring containing 1 to 4 carbon atoms and 1 to 3nitrogen or oxygen atoms and is optionally substituted with hydroxyl,protected hydroxyl, alkyloxy, amino, protected amino, alkylamino,aminoalkyl, or halogen;

[0087] R₁₀ is hydrogen, hydroxyl, protected hydroxyl, amino, or aprotected amino group;

[0088] R₁₁ is hydrogen, hydroxyl, or protected hydroxyl;

[0089] X is absent or is O, NH, or CH₂;

[0090] Y is —CHR₁₇, O, C═O, CR₁₇ or NR₁₇, and Z is CHR₁₈, O, C═O, CR₁₈or NR₁₈, wherein each occurrence of R₁₇ and R₁₈ is independentlyhydrogen or lower alkyl; and

[0091] pharmaceutically acceptable derivatives thereof.

[0092] In certain embodiments, the present invention defines certainclasses of compounds which are of special interest. For example, oneclass of compounds of special interest includes those compounds havingthe structure of formula (I) in which X is O, and n is 1 and thecompound has the structure:

[0093] wherein R₁-R₁₁, Y and Z are as previously defined.

[0094] Another class of compounds of special interest includes compoundshaving the structure of formula (I) in which R₄ is halogen (Hal), andthe compound has the structure:

[0095] wherein R₁-R₃, R₅-R₁₁, X, Y, Z and n are as previously defined,and wherein Hal is a halogen selected from fluorine, bromine, chlorineand iodine.

[0096] Another class of compounds of special interest includes compoundshaving the structure of formula (I) in which Y and Z together represent—CH═CH—, and the compound has the structure:

[0097] wherein R₁-R₁₁, X and n are as previously defined.

[0098] Another class of compounds of special interest includes compoundshaving the structure of formula (I) in which R₁ and R₂ are each methyl,and R₃ is hydrogen and the compound has the structure:

[0099] wherein R₄-R₁₁, n, X, Y and Z are as previously defined.

[0100] Another class of compounds of special interest includes compoundshaving the structure of formula (I) in which R₉ is NR₁₂R₁₃, and thecompound has the structure:

[0101] wherein R₁-R₁₃, n, X, Y and Z are as previously defined,

[0102] and R₁₃ and R₈ may additionally, when taken together, form asaturated or unsaturated cyclic ring containing 1 to 4 carbon atoms and1 to 3 nitrogen or oxygen atoms and is optionally substituted withhydroxyl, protected hydroxyl, alkyloxy, amino, protected amino,alkylamino, aminoalkyl, and halogen.

[0103] Another class of compounds of special interest includes compoundshaving the structure of formula (I) in which R₉ is OR₁₂, and thecompound has the structure:

[0104] wherein R₁-R₁₂, n, X, Y and Z are as previously defined.

[0105] Another class of compounds of special interest includes compoundshaving the structure of formula (I) in which R₉ is —X₁(CH₂)_(p)X₂R₁₄,and the compound has the structure:

[0106] wherein R₁-R₁₁, R₁₄, X, Y, Z, n, X₁, X₂ and p are as definedabove.

[0107] The following structures illustrate several exemplary types ofcompounds of these classes. Additional compounds are described in theExemplification herein. Other compounds of the invention will be readilyapparent to the reader:

[0108] A number of important subclasses of each of the foregoing classesdeserve separate mention; these subclasses include subclasses of theforegoing classes in which:

[0109] i) R₁ is hydrogen, aryl or lower alkyl;

[0110] ii) R₁ is hydrogen, phenyl, methyl or ethyl;

[0111] iii) R₁ is methyl;

[0112] iv) R₂ is hydrogen, halogen or lower alkyl;

[0113] v) R₂ is hydrogen, F, methyl or ethyl;

[0114] vi) R₂ is methyl;

[0115] vii) R₃ is hydrogen;

[0116] viii) R₁ and R₂ are each methyl and R₃ is hydrogen;

[0117] ix) R₁ and R₂, taken together, form a 5- to 6-membered cycloalkylmoiety;

[0118] x) R₁ and R₃, taken together, form a 5- to 6-membered cycloalkylmoiety;

[0119] xi) R₄ is a halogen selected from fluorine, chlorine, bromine,and iodine;

[0120] xii) R₄ is a hydrogen;

[0121] xiii) R₄ is fluorine;

[0122] xiv) R₅ is a protecting group, hydrogen or a prodrug moiety;

[0123] xv) R₅ is an oxygen protecting group;

[0124] xvi) R₅ is an oxygen protecting group selected from methyl ether,substituted methyl ether, substituted ethyl ether, substituted benzylether, silyl ether, ester, carbonate, cyclic acetal and ketal;

[0125] xvii) R₆ is hydrogen, hydroxyl or protected hydroxyl;

[0126] xviii) R₆ is protected hydroxyl and the protecting group is anoxygen protecting group;

[0127] xix) R₆ is protected hydroxyl and the protecting group is anoxygen protecting group selected from methyl ether, substituted methylether, substituted ethyl ether, substituted benzyl ether, silyl ether,ester, carbonate, cyclic acetal and ketal;

[0128] xx) R₆ is protected hydroxyl and the protecting group is aprodrug moiety;

[0129] xxi) n is 1;

[0130] xxii) R₇ is hydrogen;

[0131] xxiii) R₇ is hydroxyl;

[0132] xxiv) R₇ is protected hydroxyl and the protecting group is anoxygen protecting group;

[0133] xxv) R₇ is a protected hydroxyl and the protecting group is anoxygen protecting group selected from methyl ether, substituted methylether, substituted ethyl ether, substituted benzyl ether, silyl ether,ester, carbonate, cyclic acetal and ketal;

[0134] xxvi) R₇ is protected hydroxyl and the protecting group is aprodrug moiety;

[0135] xxvii) Y and Z together represent —CH═CH—;

[0136] xxviii) Y and Z together represent trans —CH═CH—;

[0137] xxix) Y and Z together represent —CR₁₇═CR₁₈—;

[0138] xxx) Y and Z together represent trans CR₁₇═CR₁₈—;

[0139] xxxi) Y and Z together are an epoxide;

[0140] xxxii) Y and Z together are an aziridine;

[0141] xxxiii) Y and Z together are cyclopropyl;

[0142] xxxiv) Y and Z together are —CH₂—CH₂—;

[0143] xxxv) Z is O;

[0144] xxxvi) Y is O;

[0145] xxxvii) Z is C═O and Y is CHR₁₇;

[0146] xxxviii) Z is NR₁₈ and Y is CHR₁₇;

[0147] xxxix) Z is CHR₁₈ and Y is C═O;

[0148] xl) Z is CHR₁₈ and Y is NR₁₇;

[0149] xli) X is O or NH;

[0150] xlii) R₈ is hydrogen;

[0151] xliii) R₈ is halogen, hydroxyl, protected hydroxyl, alkyloxy, orlower alkyl ptionally substituted when one or more hydroxyl or protectedhydroxyl groups;

[0152] xliv) R₉ is hydrogen;

[0153] xlv) R₉ is OR₁₂, wherein R₁₂ is methyl, ethyl, propyl, isopropyl,butyl, —CH₂COOMe, Bn, PMB (MPM), 3, 4-CIBn, or R₉ is

[0154] xlvi) R₉ is NR₁₂R₁₃, wherein R₁₂ is methyl, ethyl, propyl,isopropyl, or butyl, optionally substituted with one or more occurrencesof hydroxyl or protected hydroxyl, and R₁₃ is hydrogen or lower alkyl,or NR₁₂R₁₃ together represents a 5- or 6-membered heterocyclic moiety;

[0155] xlvii) R₉ is O(CH₂)_(p)X₂R₁₄, wherein X₂R₁₄ together representN₃, NMe₂, NHAc, NHSO₂Me, NHCONHMe, NHCONHPh, morpholine, imidazole,aminopyridine, or any one of:

[0156] xlviii) R₁₀ is hydroxyl or protected hydroxyl;

[0157] xlix) R₁₀ is hydroxyl; and/or

[0158] l) R₁₁ is hydrogen.

[0159] As the reader will appreciate, compounds of particular interestinclude, among others, those which share the attributes of one or moreof the foregoing subclasses. Some of those subclasses are illustrated bythe following sorts of compounds:

[0160] I) Compounds of the formula (and pharmaceutically acceptablederivatives thereof):

[0161] wherein R₅-R₈, R₁₀-R₁₃ are as defined above and in subclassesherein.

[0162] II) Compounds of the formula (and pharmaceutically acceptablederivatives thereof):

[0163] wherein R₅-R₈, R₁₀-R₁₃ are as defined above and in subclassesherein.

[0164] III) Compounds of the formula (and pharmaceutically acceptablederivatives thereof):

[0165] wherein R₅-R₈, R₁₀ and R₁₂ are as defined above and in subclassesherein.

[0166] IV) Compounds of the formula (and pharmaceutically acceptablederivatives thereof):

[0167] wherein R₅-R₈, R₁₀ and R₁₂ are as defined above and in subclassesherein.

[0168] V) Compounds of the formula (and pharmaceutically acceptablederivatives thereof):

[0169] wherein R₅-R₈, R₁₀, R₁₄, X₁, X₂ and p are as defined above and insubclasses herein.

[0170] VI) Compounds of the formula (and pharmaceutically acceptablederivatives thereof):

[0171] wherein R₅-R₈, R₁₀, R₁₄, X₁, X₂ and p are as defined above and insubclasses herein.

[0172] It will also be appreciated that for each of the subgroups I-VIdescribed above, a variety of other subclasses are of special interest,including, but not limited to those classes described above i)-l) andclasses, subclasses and species of compounds described above and in theexamples herein.

[0173] Some of the foregoing compounds can comprise one or moreasymmetric centers, and thus can exist in various isomeric forms, e.g.,stereoisomers and/or diastereomers. Thus, inventive compounds andpharmaceutical compositions thereof may be in the form of an individualenantiomer, diastereomer or geometric isomer, or may be in the form of amixture of stereoisomers. In certain embodiments, the compounds of theinvention are enantiopure compounds. In certain other embodiments,mixtures of stereoisomers or diastereomers are provided.

[0174] Furthermore, certain compounds, as described herein may have oneor more double bonds that can exist as either the Z or E isomer, unlessotherwise indicated. The invention additionally encompasses thecompounds as individual isomers substantially free of other isomers andalternatively, as mixtures of various isomers, e.g., racemic mixtures ofstereoisomers. In addition to the above-mentioned compounds per se, thisinvention also encompasses pharmaceutically acceptable derivatives ofthese compounds and compositions comprising one or more compounds of theinvention and one or more pharmaceutically acceptable excipients oradditives.

[0175] Compounds of the invention may be prepared by crystallization ofcompound of formula (I) under different conditions and may exist as oneor a combination of polymorphs of compound of general formula (I)forming part of this invention. For example, different polymorphs may beidentified and/or prepared using different solvents, or differentmixtures of solvents for recrystallization; by performingcrystallizations at different temperatures; or by using various modes ofcooling, ranging from very fast to very slow cooling duringcrystallizations. Polymorphs may also be obtained by heating or meltingthe compound followed by gradual or fast cooling. The presence ofpolymorphs may be determined by solid probe NMR spectroscopy, IRspectroscopy, differential scanning calorimetry, powder X-raydiffractogram and/or other techniques. Thus, the present inventionencompasses inventive compounds, their derivatives, their tautomericforms, their stereoisomers, their polymorphs, their pharmaceuticallyacceptable salts their pharmaceutically acceptable solvates andpharmaceutically acceptable compositions containing them.

[0176] 2) Compounds and Definitions

[0177] As discussed above, this invention provides novel compounds witha range of biological properties. Compounds of this invention havebiological activities relevant for the treatment of inflammatory andimmune disorders, photoaging and cancer. In certain embodiments, thecompounds of the invention are useful for the treatment of rheumatoidarthritis, psoriasis, Multiple sclerosis, and asthma. In certain otherembodiments, the inventive compounds also find use in the prevention ofrestenosis of blood vessels subject to traumas such as angioplasty andstenting.

[0178] Compounds of this invention include those specifically set forthabove and described herein, and are illustrated in part by the variousclasses, subgenera and species disclosed elsewhere herein.

[0179] Additionally, the present invention provides pharmaceuticallyacceptable derivatives of the inventive compounds, and methods oftreating a subject using these compounds, pharmaceutical compositionsthereof, or either of these in combination with one or more additionaltherapeutic agents. The phrase, “pharmaceutically acceptablederivative”, as used herein, denotes any pharmaceutically acceptablesalt, ester, or salt of such ester, of such compound, or any otheradduct or derivative which, upon administration to a patient, is capableof providing (directly or indirectly) a compound as otherwise describedherein, or a metabolite or residue thereof. Pharmaceutically acceptablederivatives thus include among others pro-drugs. A pro-drug is aderivative of a compound, usually with significantly reducedpharmacological activity, which contains an additional moiety, which issusceptible to removal in vivo yielding the parent molecule as thepharmacologically active species. An example of a pro-drug is an ester,which is cleaved in vivo to yield a compound of interest. Pro-drugs of avariety of compounds, and materials and methods for derivatizing theparent compounds to create the pro-drugs, are known and may be adaptedto the present invention. Certain exemplary pharmaceutical compositionsand pharmaceutically acceptable derivatives will be discussed in moredetail herein below.

[0180] Certain compounds of the present invention, and definitions ofspecific functional groups are also described in more detail below. Forpurposes of this invention, the chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 75^(th) Ed., inside cover, andspecific functional groups are generally defined as described therein.Additionally, general principles of organic chemistry, as well asspecific functional moieties and reactivity, are described in “OrganicChemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999,the entire contents of which are incorporated herein by reference.Furthermore, it will be appreciated by one of ordinary skill in the artthat the synthetic methods, as described herein, utilize a variety ofprotecting groups. By the term “protecting group”, has used herein, itis meant that a particular functional moiety, e.g., O, S, or N, istemporarily blocked so that a reaction can be carried out selectively atanother reactive site in a multifunctional compound. In preferredembodiments, a protecting group reacts selectively in good yield to givea protected substrate that is stable to the projected reactions; theprotecting group must be selectively removed in good yield by readilyavailable, preferably nontoxic reagents that do not attack the otherfunctional groups; the protecting group forms an easily separablederivative (more preferably without the generation of new stereogeniccenters); and the protecting group has a minimum of additionalfunctionality to avoid further sites of reaction. As detailed herein,oxygen, sulfur, nitrogen and carbon protecting groups may be utilized.For example, in certain embodiments, as detailed herein, certainexemplary oxygen protecting groups are utilized. These oxygen protectinggroups include, but are not limited to methyl ethers, substituted methylethers (e.g., MOM (methoxymethyl ether), MTM (methylthiomethyl ether),BOM (benzyloxymethyl ether), PMBM or MPM (p-methoxybenzyloxymethylether), to name a few), substituted ethyl ethers, substituted benzylethers, silyl ethers (e.g., TMS (trimethylsilyl ether), TES(triethylsilylether), TIPS (triisopropylsilyl ether), TBDMS(t-butyldimethylsilyl ether), tribenzyl silyl ether, TBDPS(t-butyldiphenyl silyl ether), to name a few), esters (e.g., formate,acetate, benzoate (Bz), trifluoroacetate, dichloroacetate, to name afew), carbonates, cyclic acetals and ketals. In certain other exemplaryembodiments, nitrogen protecting groups are utilized. These nitrogenprotecting groups include, but are not limited to, carbamates (includingmethyl, ethyl and substituted ethyl carbamates (e.g., Troc), to name afew) amides, cyclic imide derivatives, N-Alkyl and N-Aryl amines, iminederivatives, and enamine derivatives, to name a few. Certain otherexemplary protecting groups are detailed herein, however, it will beappreciated that the present invention is not intended to be limited tothese protecting groups; rather, a variety of additional equivalentprotecting groups can be readily identified using the above criteria andutilized in the present invention. Additionally, a variety of protectinggroups are described in “Protective Groups in Organic Synthesis” ThirdEd. Greene, T. W. and Wuts, P. G., Eds., John Wiley & Sons, New York:1999, the entire contents of which are hereby incorporated by reference.

[0181] It will be appreciated that the compounds, as described herein,may be substituted with any number of substituents or functionalmoieties. In general, the term “substituted” whether preceded by theterm “optionally” or not, and substituents contained in formulas of thisinvention, refer to the replacement of hydrogen radicals in a givenstructure with the radical of a specified substituent. When more thanone position in any given structure may be substituted with more thanone substituent selected from a specified group, the substituent may beeither the same or different at every position. As used herein, the term“substituted” is contemplated to include all permissible substituents oforganic compounds. In a broad aspect, the permissible substituentsinclude acyclic and cyclic, branched and unbranched, carbocyclic andheterocyclic, aromatic and nonaromatic substituents of organiccompounds. For purposes of this invention, heteroatoms such as nitrogenmay have hydrogen substituents and/or any permissible substituents oforganic compounds described herein which satisfy the valencies of theheteroatoms. Furthermore, this invention is not intended to be limitedin any manner by the permissible substituents of organic compounds.Combinations of substituents and variables envisioned by this inventionare preferably those that result in the formation of stable compoundsuseful in the treatment, for example of inflammatory and proliferativedisorders, including, but not limited to rheumatoid arthritis,psoriasis, asthma and cancer. The term “stable”, as used herein,preferably refers to compounds which possess stability sufficient toallow manufacture and which maintain the integrity of the compound for asufficient period of time to be detected and preferably for a sufficientperiod of time to be useful for the purposes detailed herein.

[0182] The term “aliphatic”, as used herein, includes both saturated andunsaturated, straight chain (i.e., unbranched) or branched aliphatichydrocarbons, which are optionally substituted with one or morefunctional groups. As will be appreciated by one of ordinary skill inthe art, “aliphatic” is intended herein to include, but is not limitedto, alkyl, alkenyl, alkynyl moieties. Thus, as used herein, the term“alkyl” includes straight and branched alkyl groups. An analogousconvention applies to other generic terms such as “alkenyl”, “alkynyl”and the like. Furthermore, as used herein, the terms “alkyl”, “alkenyl”,“alkynyl” and the like encompass both substituted and unsubstitutedgroups. In certain embodiments, as used herein, “lower alkyl” is used toindicate those alkyl groups (cyclic, acyclic, substituted,unsubstituted, branched or unbranched) having 1-6 carbon atoms.

[0183] In certain embodiments, the alkyl, alkenyl and alkynyl groupsemployed in the invention contain 1-20 aliphatic carbon atoms. Incertain other embodiments, the alkyl, alkenyl, and alkynyl groupsemployed in the invention contain 1-10 aliphatic carbon atoms. In yetother embodiments, the alkyl, alkenyl, and alkynyl groups employed inthe invention contain 1-8 aliphatic carbon atoms. In still otherembodiments, the alkyl, alkenyl, and alkynyl groups employed in theinvention contain 1-6 aliphatic carbon atoms. In yet other embodiments,the alkyl, alkenyl, and alkynyl groups employed in the invention contain1-4 carbon atoms. Illustrative aliphatic groups thus include, but arenot limited to, for example, methyl, ethyl, n-propyl, isopropyl, allyl,n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, sec-pentyl,isopentyl, tert-pentyl, n-hexyl, sec-hexyl, moieties and the like, whichagain, may bear one or more substituents. Alkenyl groups include, butare not limited to, for example, ethenyl, propenyl, butenyl,1-methyl-2-buten-1-yl, and the like. Representative alkynyl groupsinclude, but are not limited to, ethynyl, 2-propynyl (propargyl),1-propynyl and the like.

[0184] The term “alicyclic”, as used herein, refers to compounds whichcombine the properties of aliphatic and cyclic compounds and include butare not limited to cyclic, or polycyclic aliphatic hydrocarbons andbridged cycloalkyl compounds, which are optionally substituted with oneor more functional groups. As will be appreciated by one of ordinaryskill in the art, “alicyclic” is intended herein to include, but is notlimited to, cycloalkyl, cycloalkenyl, and cycloalkynyl moieties, whichare optionally substituted with one or more functional groups.Illustrative alicyclic groups thus include, but are not limited to, forexample, cyclopropyl, —CH₂-cyclopropyl, cyclobutyl, —CH₂-cyclobutyl,cyclopentyl, —CH₂-cyclopentyl-n, cyclohexyl, —CH₂-cyclohexyl,cyclohexenylethyl, cyclohexanylethyl, norborbyl moieties and the like,which again, may bear one or more substituents.

[0185] The term “alkoxy” (or “alkyloxy”), or “thioalkyl” as used hereinrefers to an alkyl group, as previously defined, attached to the parentmolecular moiety through an oxygen atom or through a sulfur atom. Incertain embodiments, the alkyl group contains 1-20 aliphatic carbonatoms. In certain other embodiments, the alkyl group contains 1-10aliphatic carbon atoms. In yet other embodiments, the alkyl, alkenyl,and alkynyl groups employed in the invention contain 1-8 aliphaticcarbon atoms. In still other embodiments, the alkyl group contains 1-6aliphatic carbon atoms. In yet other embodiments, the alkyl groupcontains 1-4 aliphatic carbon atoms. Examples of alkoxy, include but arenot limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy,tert-butoxy, neopentoxy and n-hexoxy. Examples of thioalkyl include, butare not limited to, methylthio, ethylthio, propylthio, isopropylthio,n-butylthio, and the like.

[0186] The term “alkylamino” refers to a group having the structure—NHR′wherein R′ is alkyl, as defined herein. The term “aminoalkyl”refers to a group having the structure NH₂R′—, wherein R′ is alkyl, asdefined herein. In certain embodiments, the alkyl group contains 1-20aliphatic carbon atoms. In certain other embodiments, the alkyl groupcontains 1-10 aliphatic carbon atoms. In yet other embodiments, thealkyl, alkenyl, and alkynyl groups employed in the invention contain 1-8aliphatic carbon atoms. In still other embodiments, the alkyl groupcontains 1-6 aliphatic carbon atoms. In yet other embodiments, the alkylgroup contains 1-4 aliphatic carbon atoms. Examples of alkylaminoinclude, but are not limited to, methylamino, ethylamino,iso-propylamino and the like.

[0187] Some examples of substituents of the above-described aliphatic(and other) moieties of compounds of the invention include, but are notlimited to aliphatic; heteroaliphatic; aryl; heteroaryl; alkylaryl;alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy; heteroaryloxy;alkylthio; arylthio; heteroalkylthio; heteroarylthio; F; Cl; Br; I; —OH;—NO₂; —CN; —CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH; —CH₂CH₂OH; —CH₂NH₂;—CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x);—OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂; —S(O)₂R_(x); —NR_(x)(CO)R_(x)wherein each occurrence of R_(x) independently includes, but is notlimited to, aliphatic, heteroaliphatic, aryl, heteroaryl, alkylaryl, oralkylheteroaryl, wherein any of the aliphatic, heteroaliphatic,alkylaryl, or alkylheteroaryl substituents described above and hereinmay be substituted or unsubstituted, branched or unbranched, cyclic oracyclic, and wherein any of the aryl or heteroaryl substituentsdescribed above and herein may be substituted or unsubstituted.Additional examples of generally applicable substituents are illustratedby the specific embodiments shown in the Examples that are describedherein.

[0188] In general, the terms “aryl” and “heteroaryl”, as used herein,refer to stable mono- or polycyclic, heterocyclic, polycyclic, andpolyheterocyclic unsaturated moieties having preferably 3-14 carbonatoms, each of which may be substituted or unsubstituted. It will alsobe appreciated that aryl and heteroaryl moieties, as defined herein maybe attached via an aliphatic, alicyclic, heteroaliphatic,heteroalicyclic, alkyl or heteroalkyl moiety and thus also include-(aliphatic)aryl, -(heteroaliphatic)aryl, -(aliphatic)heteroaryl,-(heteroaliphatic)heteroaryl, -(alkyl)aryl, -(heteroalkyl)aryl,-(heteroalkyl)aryl, and -(heteroalkyl)heteroaryl moieties. Thus, as usedherein, the phrases “aryl or heteroaryl” and “aryl, heteroaryl,-(aliphatic)aryl, -(heteroaliphatic)aryl, -(aliphatic)heteroaryl,-(heteroaliphatic)heteroaryl, -(alkyl)aryl, -(heteroalkyl)aryl,-(heteroalkyl)aryl, and -(heteroalkyl)heteroaryl” are interchangeable.Substituents include, but are not limited to, any of the previouslymentioned substitutents, i.e., the substituents recited for aliphaticmoieties, or for other moieties as disclosed herein, resulting in theformation of a stable compound. In certain embodiments of the presentinvention, “aryl” refers to a mono- or bicyclic carbocyclic ring systemhaving one or two aromatic rings including, but not limited to, phenyl,naphthyl, tetrahydronaphthyl, indanyl, indenyl and the like. In certainembodiments of the present invention, the term “heteroaryl”, as usedherein, refers to a cyclic aromatic radical having from five to ten ringatoms of which one ring atom is selected from S, O and N; zero, one ortwo ring atoms are additional heteroatoms independently selected from S,O and N; and the remaining ring atoms are carbon, the radical beingjoined to the rest of the molecule via any of the ring atoms, such as,for example, pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl,imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl,thiophenyl, furanyl, quinolinyl, isoquinolinyl, and the like.

[0189] It will be appreciated that aryl and heteroaryl groups (includingbicyclic aryl groups) can be unsubstituted or substituted, whereinsubstitution includes replacement of one, two or three of the hydrogenatoms thereon independently with any one or more of the followingmoieties including, but not limited to: aliphatic; heteroaliphatic;aryl; heteroaryl; alkylaryl; alkylheteroaryl; alkoxy; aryloxy;heteroalkoxy; heteroaryloxy; alkylthio; arylthio; heteroalkylthio;heteroarylthio; F; Cl; Br; I; —OH; —NO₂; —CN; —CF₃; —CH₂CF₃; —CHCl₂;—CH₂OH; —CH₂CH₂OH; —CH₂NH₂; —CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x));—CON(R_(x))₂; —OC(O)R_(x); —OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂;—S(O)₂R_(x); —NR_(x)(CO)R_(x) wherein each occurrence of R_(x)independently includes, but is not limited to, aliphatic,heteroaliphatic, aryl, heteroaryl, alkylaryl, or alkylheteroaryl,wherein any of the aliphatic, heteroaliphatic, alkylaryl, oralkylheteroaryl substituents described above and herein may besubstituted or unsubstituted, branched or unbranched, cyclic or acyclic,and wherein any of the aryl or heteroaryl substituents described aboveand herein may be substituted or unsubstituted. Additional examples ofgenerally applicable substituents are illustrated by the specificembodiments shown in the Examples that are described herein.

[0190] The term “cycloalkyl”, as used herein, refers specifically togroups having three to seven, preferably three to ten carbon atoms.Suitable cycloalkyls include, but are not limited to cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like, which, asin the case of aliphatic, heteroaliphatic or heterocyclic moieties, mayoptionally be substituted with substituents including, but not limitedto aliphatic; heteroaliphatic; aryl; heteroaryl; alkylaryl;alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy; heteroaryloxy;alkylthio; arylthio; heteroalkylthio; heteroarylthio; F; Cl; Br; I; —OH;—NO₂; —CN; —CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH; —CH₂CH₂OH; —CH₂NH₂;—CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x);—OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂; —S(O)₂R_(x); —NR_(x)(CO)R_(x)wherein each occurrence of R_(x) independently includes, but is notlimited to, aliphatic, heteroaliphatic, aryl, heteroaryl, alkylaryl, oralkylheteroaryl, wherein any of the aliphatic, heteroaliphatic,alkylaryl, or alkylheteroaryl substituents described above and hereinmay be substituted or unsubstituted, branched or unbranched, cyclic oracyclic, and wherein any of the aryl or heteroaryl substituentsdescribed above and herein may be substituted or unsubstituted.Additional examples of generally applicable substituents are illustratedby the specific embodiments shown in the Examples that are describedherein.

[0191] The term “heteroaliphatic”, as used herein, refers to aliphaticmoieties in which one or more carbon atoms in the main chain have beensubstituted with a heteroatom. Thus, a heteroaliphatic group refers toan aliphatic chain which contains one or more oxygen, sulfur, nitrogen,phosphorus or silicon atoms, e.g., in place of carbon atoms.Heteroaliphatic moieties may be branched or linear unbranched. Incertain embodiments, heteroaliphatic moieties are substituted byindependent replacement of one or more of the hydrogen atoms thereonwith one or more moieties including, but not limited to aliphatic;alicyclic; heteroaliphatic; heteroalicyclic; aryl; heteroaryl;alkylaryl; alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy;heteroaryloxy; alkylthio; arylthio; heteroalkylthio; heteroarylthio; F;Cl; Br; I; —OH; —NO₂; —CN; —CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH; —CH₂CH₂OH;—CH₂NH₂; —CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x);—OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂; —S(O)₂R_(x); —NR_(x)(CO)R_(x)wherein each occurrence of R_(x) independently includes, but is notlimited to, aliphatic, alicyclic, heteroaliphatic, heteroalicyclic,aryl, heteroaryl, alkylaryl, or alkylheteroaryl, wherein any of thealiphatic, alicyclic, heteroaliphatic, heteroalicyclic, alkylaryl, oralkylheteroaryl substituents described above and herein may besubstituted or unsubstituted, branched or unbranched, cyclic or acyclic,and wherein any of the aryl or heteroaryl substituents described aboveand herein may be substituted or unsubstituted. Additional examples ofgenerally applicable substituents are illustrated by the specificembodiments shown in the Examples that are described herein.

[0192] The term “heteroalicyclic”, as used herein, refers to compoundswhich combine the properties of heteroaliphatic and cyclic compounds andinclude but are not limited to saturated and unsaturated mono- orpolycyclic heterocycles such as morpholino, pyrrolidinyl, furanyl,thiofuranyl, pyrrolyl etc., which are optionally substituted with one ormore functional groups, as defined herein.

[0193] Additionally, it will be appreciated that any of the alicyclic orheteroalicyclic moieties described above and herein may comprise an arylor heteroaryl moiety fused thereto. Additional examples of generallyapplicable substituents are illustrated by the specific embodimentsshown in the Examples that are described herein.

[0194] The terms “halo” and “halogen” as used herein refer to an atomselected from fluorine, chlorine, bromine and iodine.

[0195] The term “haloalkyl” denotes an alkyl group, as defined above,having one, two, or three halogen atoms attached thereto and isexemplified by such groups as chloromethyl, bromoethyl, trifluoromethyl,and the like.

[0196] The term “heterocycloalkyl” or “heterocycle”, as used herein,refers to a non-aromatic 5-, 6- or 7- membered ring or a polycyclicgroup, including, but not limited to a bi- or tri-cyclic groupcomprising fused six-membered rings having between one and threeheteroatoms independently selected from oxygen, sulfur and nitrogen,wherein (i) each 5-membered ring has 0 to 1 double bonds and each6-membered ring has 0 to 2 double bonds, (ii) the nitrogen and sulfurheteroatoms may be optionally be oxidized, (iii) the nitrogen heteroatommay optionally be quaternized, and (iv) any of the above heterocyclicrings may be fused to an aryl or heteroaryl ring. Representativeheterocycles include, but are not limited to, pyrrolidinyl, pyrazolinyl,pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl,oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl,isothiazolidinyl, and tetrahydrofuryl. In certain embodiments, a“substituted heterocycloalkyl or heterocycle” group is utilized and asused herein, refers to a heterocycloalkyl or heterocycle group, asdefined above, substituted by the independent replacement of one, two orthree of the hydrogen atoms thereon with but are not limited toaliphatic; heteroaliphatic; aryl; heteroaryl; alkylaryl;alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy; heteroaryloxy;alkylthio; arylthio; heteroalkylthio; heteroarylthio; F; Cl; Br; I; —OH;—NO₂; —CN; —CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH; —CH₂CH₂OH; —CH₂NH₂;—CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x);—OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂; —S(O)₂R_(x);—NR_(x)(CO)R_(x)wherein each occurrence of R_(x) independently includes,but is not limited to, aliphatic, heteroaliphatic, aryl, heteroaryl,alkylaryl, or alkylheteroaryl, wherein any of the aliphatic,heteroaliphatic, alkylaryl, or alkylheteroaryl substituents describedabove and herein may be substituted or unsubstituted, branched orunbranched, cyclic or acyclic, and wherein any of the aryl or heteroarylsubstitutents described above and herein may be substituted orunsubstituted. Additional examples or generally applicable substituentsare illustrated by the specific embodiments shown in the Examples, whichare described herein.

[0197] As used herein, the terms “aliphatic”, “heteroaliphatic”,“alkyl”, “alkenyl”, “alkynyl”, “heteroalkyl”, “heteroalkenyl”,“heteroalkynyl”, and the like encompass substituted and unsubstituted,saturated and unsaturated, and linear and branched groups. Similarly,the terms “alicyclic”, “heteroalicyclic”, “heterocycloalkyl”,“heterocycle” and the like encompass substituted and unsubstituted, andsaturated and unsaturated groups. Additionally, the terms “cycloalkyl”,“cycloalkenyl”, “cycloalkynyl”, “heterocycloalkyl”,“heterocycloalkenyl”, “heterocycloalkynyl”, “aryl”, “heteroaryl” and thelike encompass both substituted and unsubstituted groups.

[0198] 3) Synthetic Methodology

[0199] As described above, the present invention provides novelmacrocycles having formula (I) a described above and in certain classesand subclasses herein. An overview of an exemplary synthesis of theinventive compounds is provided below, as detailed in Schemes 1-7, andin the Exemplification herein. It will be appreciated that the methodsas described herein can be applied to each of the compounds as disclosedherein and equivalents thereof. Additionally, the reagents and startingmaterials are well known to those skilled in the art. Although thefollowing schemes describe certain exemplary compounds, it will beappreciated that the use of alternate starting materials will yieldother analogs of the invention. For example, compounds are describedbelow where X is O; however, it will be appreciated that alternatestarting materials and/or intermediates can be utilized to generatecompounds where X is NH, N-alkyl, CH₂, etc.

[0200] In general, compounds as provided herein especially modificationwhere Y and Z together as CH═CH or CH₂CH₂, are prepared from assembly ofthese three segments in two different orders depending on position ofmodifications on the ring, as depicted below.

[0201] For R₄ modifications, a third route was use to incorporate R₄ asdepicted below:

[0202] For compounds with Y and Z are heteroatom such N, O or CO, adifferent set of reaction condition was used to form these bonds inplace of a C—C bond formation. Certain analogs were prepared usingvariation of these methods shown above.

[0203] For R₉ analogs, compounds as provided herein, are prepared from ageneral advance intermediate in additional to the general methodsdescribed above, as depicted below (2):

[0204] In certain embodiments, this general advance intermediate can besynthesized from two components, an aromatic component, the synthesis ofwhich is depicted in Scheme 1 and is described in more detail inexamples herein, and a protected diol component, the synthesis of whichis depicted in Scheme 2 and is described in more detail in examplesherein. As depicted in Scheme 3, and as described in more detail inexamples herein, these two components are coupled, and subsequentreduction to generate the double bond occurs. Finally, macrocyclizationis effected to generate the macrolactone intermediate.

[0205] As depicted in Scheme 4, and as described in the examples herein,an alternate route to the protected diol intermediate provides facileaccess to compounds where R₄ is halogen. Coupling of this intermediatewith the aromatic component described above and herein, providesadditional structures where R₄ is halogen, or, as depicted, F.

[0206] It will be appreciated that once the core intermediate structuresare constructed a variety of other analogues can be generated. In butone example, C14-O analogues are provided (R₉ as described herein). Forexample, Scheme 5 depicts the synthesis of these analogues using aMitsunobu reaction to functionalize the C14 hydroxyl moiety.

[0207] Alternatively, as depicted in Scheme 6, the hydroxylfunctionality in the advance intermediate can be replaced with an aminefunctionality. This amine can be further substituted (e.g., with methylgroups, as depicted in Scheme 6) with a variety of functional groups asdescribed herein, using methods available to one of ordinary skill inthe art.

[0208] Alternatively, as depicted in Schemes 7 and 9, the aminefunctionality may be introduced earlier in the synthesis. This amine canbe further substituted (e.g., with methyl or ethyl groups, as depictedin Schemes 7 and 9) with a variety of functional groups as describedherein, using methods available to one of ordinary skill in the art. Asynthesis of acyclic intermediate 20 is depicted in Scheme 8.

[0209] For special fused ring systems on the aromatic component, adifferent aromatic segment is used in the place of the phenol. Whilesynthesis of the aromatic fragment required special synthetictechniques, the overall flow remained, as depicted below (Scheme 10)

[0210] 4) Research Uses, Formulation and Administration

[0211] According to the present invention, the inventive compounds maybe assayed in any of the available assays known in the art foridentifying compounds having antiangiogenic activity, anti-inflammatoryactivity, protein kinase inhibitory activity, NF-κB activationinhibitory activity activity and AP-1 activation inhibitory activity.For example, the assay may be cellular or non-cellular, in vivo or invitro, high- or low-throughput format, etc.

[0212] Thus, in one aspect, compounds of this invention which are ofparticular interest include those which:

[0213] exhibit activity as inhibitors of NF-κB activation, AP-1activation and protein kinases (e.g., MEKK1, MEK1, VEGFr, PDGFr);

[0214] exhibit activity as inhibitors of production of pro-inflammatoryand/or immunologic cytokines (e.g., TNFα, IL-1, IL-6, IL-8, IL-2);

[0215] exhibit an antiproliferative or an antiangiogenic effect on solidtumors;

[0216] exhibit an anti-inflammatory effect on suitable cell linesmaintained in vitro, or in animal studies using a scientificallyacceptable model;

[0217] are useful for the treatment of photoaging-relateddisorders/conditions; and/or

[0218] exhibit a favorable therapeutic profile (e.g., safety, efficacy,and stability).

[0219] As discussed above, certain of the compounds as described hereinexhibit activity generally as inhibitors of NF-κB activation, AP-1activation and protein kinases. More specifically, compounds of theinvention demonstrate immnosuppressive activity and thus the inventionfurther provides a method for treating an inflammatory disorder orautoimmune disorders. Certain of the compounds as described herein alsoact as inhibitors of tumor growth and angiogenesis. The method involvesthe administration of a therapeutically effective amount of the compoundor pharmaceutically acceptable derivative thereof to a subject(including, but not limited to a human or animal) in need of it. Incertain embodiments, the inventive compounds as useful for the treatmentof sepsis, glomerulonephropathy, rheumatoid arthritis (includingankylosing spondylitis), psoriatic arthritis, osteoarthritis,osteoporosis, allergic rhinitis, ocular inflammation, inflammatory boweldisease (crohn's disease and ulcerative colitis), multiple sclerosis,atopic dermatitis, psoriasis, asthma, inflammatory pulmonary disease,hepatitis, autoimmune disorders, systemic lupus erthematosus, allograftrejection/graft versus host disease, diabetes, AIDS, solid tumorcancers, leukemia, lymphomas, non-hodgkin's B-cell lymphomas, chronicallymphocytic leukemia (CLL), multiple myeloma, eczema, urticaria,myasthenia gravis, idiopathic thrombocytopenia purpura, cardiovasculardisease (e.g., myocardial infarction, atherosclerosis), hepatitis,glomerulonephropathy, productive nephritis, adenovirus,diseases/disorders of the central nervous system (e.g., stroke,Alzheimer's disease, epilepsy) and for the treatment of the symptoms ofmalaria, to name a few.

[0220] In certain other embodiments, compounds of the invention areuseful for reducing photodamage, and thus, the invention furtherprovides a method for treating photoaging-related disorders/conditions.In certain exemplary embodiments, compounds of the invention are usefulfor the treatment and/or prevention of skin coarseness, wrinkling,mottled pigmentation, sallowness, laxity, telangiectasia, lentigines,purpura and easy bruising, atrophy, fibrotic depigmented areas, andultimately premalignant and malignant neoplasms. In certain otherexemplary embodiments, compounds of the invention are useful for thetreatment and/or prevention of wrinkles and/or skin cancer.

[0221] Pharmaceutical Compositions

[0222] As discussed above this invention provides novel compounds thathave biological properties useful for the treatment of inflammatory andautoimmune disorders, photoaging and cancer. The inventive compoundsalso find use in the prevention of restenosis of blood vessels subjectto traumas such as angioplasty and stenting. Accordingly, in anotheraspect of the present invention, pharmaceutical compositions areprovided, which comprise any one of the compounds described herein (or aprodrug, pharmaceutically acceptable salt or other pharmaceuticallyacceptable derivative thereof), and optionally comprise apharmaceutically acceptable carrier. In certain embodiments, thesecompositions optionally further comprise one or more additionaltherapeutic agents. Alternatively, a compound of this invention may beadministered to a patient in need thereof in combination with theadministration of one or more other therapeutic agents. For example,additional therapeutic agents for conjoint administration or inclusionin a pharmaceutical composition with a compound of this invention may bean immunomodulatory agent (e.g., an agent for the treatment of,rheumatoid arthritis, psoriasis, multiple sclerosis, or asthma) orantiangiogenesis agent or anticancer agent approved for the treatment ofcancer, as discussed in more detail herein, or it may be any one of anumber of agents undergoing approval in the Food and Drug Administrationthat ultimately obtain approval for the treatment of an immune disorderor cancer. It will also be appreciated that certain of the compounds ofpresent invention can exist in free form for treatment, or whereappropriate, as a pharmaceutically acceptable derivative thereof.According to the present invention, a pharmaceutically acceptablederivative includes, but is not limited to, pharmaceutically acceptablesalts, esters, salts of such esters, or a prodrug or other adduct orderivative of a compound of this invention which upon administration toa patient in need is capable of providing, directly or indirectly, acompound as otherwise described herein, or a metabolite or residuethereof.

[0223] As used herein, the term “pharmaceutically acceptable salt”refers to those salts which are, within the scope of sound medicaljudgment, suitable for use in contact with the tissues of humans andlower animals without undue toxicity, irritation, allergic response andthe like, and are commensurate with a reasonable benefit/risk ratio.Pharmaceutically acceptable salts of amines, carboxylic acids, and othertypes of compounds, are well known in the art. For example, S. M. Berge,et al. describe pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 1-19 (1977), incorporated herein byreference. The salts can be prepared in situ during the final isolationand purification of the compounds of the invention, or separately byreacting a free base or free acid function with a suitable reagent, asdescribed generally below. For example, a free base function can bereacted with a suitable acid. Furthermore, where the compounds of theinvention carry an acidic moiety, suitable pharmaceutically acceptablesalts thereof may, include metal salts such as alkali metal salts, e.g.sodium or potassium salts; and alkaline earth metal salts, e.g. calciumor magnesium salts. Examples of pharmaceutically acceptable, nontoxicacid addition salts are salts of an amino group formed with inorganicacids such as hydrochloric acid, hydrobromic acid, phosphoric acid,sulfuric acid and perchloric acid or with organic acids such as aceticacid, oxalic acid, maleic acid, tartaric acid, citric acid, succinicacid or malonic acid or by using other methods used in the art such asion exchange. Other pharmaceutically acceptable salts include adipate,alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate,borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Representative alkali or alkaline earth metal salts includesodium, lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, loweralkyl sulfonate and aryl sulfonate.

[0224] Additionally, as used herein, the term “pharmaceuticallyacceptable ester” refers to esters that hydrolyze in vivo and includethose that break down readily in the human body to leave the parentcompound or a salt thereof. Suitable ester groups include, for example,those derived from pharmaceutically acceptable aliphatic carboxylicacids, particularly alkanoic, alkenoic, cycloalkanoic and alkanedioicacids, in which each alkyl or alkenyl moeity advantageously has not morethan 6 carbon atoms. Examples of particular esters include formates,acetates, propionates, butyrates, acrylates and ethylsuccinates.

[0225] Furthermore, the term “pharmaceutically acceptable prodrugs” asused herein refers to those prodrugs of the compounds of the presentinvention which are, within the scope of sound medical judgment,suitable for use in contact with the issues of humans and lower animalswith undue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, and effective fortheir intended use, as well as the zwitterionic forms, where possible,of the compounds of the invention. The term “prodrug” refers tocompounds that are rapidly transformed in vivo to yield the parentcompound of the above formula, for example by hydrolysis in blood. Athorough discussion is provided in T. Higuchi and V. Stella, Pro-drugsas Novel Delivery Systems, Vol. 14 of the A.C.S. Symposium Series, andin Edward B. Roche, ed., Bioreversible Carriers in Drug Design, AmericanPharmaceutical Association and Pergamon Press, 1987, both of which areincorporated herein by reference.

[0226] As described above, the pharmaceutical compositions of thepresent invention additionally comprise a pharmaceutically acceptablecarrier, which, as used herein, includes any and all solvents, diluents,or other liquid vehicle, dispersion or suspension aids, surface activeagents, isotonic agents, thickening or emulsifying agents,preservatives, solid binders, lubricants and the like, as suited to theparticular dosage form desired. Remington's Pharmaceutical Sciences,Sixteenth Edition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1980)discloses various carriers used in formulating pharmaceuticalcompositions and known techniques for the preparation thereof. Exceptinsofar as any conventional carrier medium is incompatible with thecompounds of the invention, such as by producing any undesirablebiological effect or otherwise interacting in a deleterious manner withany other component(s) of the pharmaceutical composition, its use iscontemplated to be within the scope of this invention. Some examples ofmaterials which can serve as pharmaceutically acceptable carriersinclude, but are not limited to, sugars such as lactose, glucose andsucrose; starches such as corn starch and potato starch; cellulose andits derivatives such as sodium carboxymethyl cellulose, ethyl celluloseand cellulose acetate; powdered tragacanth; malt; gelatine; talc;excipients such as cocoa butter and suppository waxes; oils such aspeanut oil, cottonseed oil; safflower oil, sesame oil; olive oil; cornoil and soybean oil; glycols; such as propylene glycol; esters such asethyl oleate and ethyl laurate; agar; buffering agents such as magnesiumhydroxide and aluminum hydroxide; alginic acid; pyrogenfree water;isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffersolutions, as well as other non-toxic compatible lubricants such assodium lauryl sulfate and magnesium stearate, as well as coloringagents, releasing agents, coating agents, sweetening, flavoring andperfuming agents, preservatives and antioxidants can also be present inthe composition, according to the judgment of the formulator.

[0227] Uses and Formulations of Compounds of the Invention

[0228] As described in more detail herein, in general, the presentinvention provides compounds useful for the treatment of inflammatory orimmune disorders and the treatment of cancer, particularly solid tumors.Without wishing to be bound by any particular theory, more generally,the compounds of the invention have been shown to inhibit NF-κB activityand the identification of NF-κB as a key player in the pathogenesis ofinflammation suggest that NF-κB targeted therapeutics may be effectivein inflammatory and immune disorders (see, generally, NF-κB in Defenseand Disease, J. Clin. Investig. 2001, 107, 7). Furthermore, certaincompounds of the invention have also been shown to inhibit receptortyrosine kinase activity such as VEGFr and PDGFr in vitro, as describedin more detail herein, and are useful for the treatment of cancer,including solid tumors (see, Angiogenesis: Potentials for PharmacologicIntervention in the Treatment of Cancer, Cardiovascular Diseases, andChronic Inflammation, Pharmacological Reviews, 2000, 52, 237).

[0229] As detailed in the exemplification herein, in assays to determinethe ability of compounds to inhibit NF-κB, certain inventive compoundsexhibited IC₅₀ values less than 10 μM. In certain other embodiments,inventive compounds exhibit IC₅₀ values less than 7.5 μM. In certainembodiments, inventive compounds exhibit IC₅₀ values less than 5 μM. Incertain other embodiments, inventive compounds exhibit IC₅₀ values lessthan 2.5 μM. In certain embodiments, inventive compounds exhibit IC₅₀values less than 1 μM. In certain embodiments, inventive compoundsexhibit IC₅₀ values less than 0.75 μM. In certain embodiments, inventivecompounds exhibit IC₅₀ values less than 0.5 μM. In certain embodiments,inventive compounds exhibit IC₅₀ values less than 0.25 μM. In certainembodiments, inventive compounds exhibit IC₅₀ values less than 0.1 μM.In certain other embodiments, inventive compounds exhibit IC₅₀ valuesless than 75 nM. In certain other embodiments, inventive compoundsexhibit IC₅₀ values less than 50 nM. In certain other embodiments,inventive compounds exhibit IC₅₀ values less than 25 nM. In certainother embodiments, inventive compounds exhibit IC₅₀ values less than 10nM. In other embodiments, exemplary compounds exhibit IC₅₀ values lessthan 7.5 nM. In other embodiments, exemplary compounds exhibit IC₅₀values less than 5 nM.

[0230] In still other embodiments, certain compounds were tested fortheir ability to inhibit the growth of tumor cell lines in vitro.Certain of these compounds exhibited IC₅₀ values less than 10 μM. Incertain other embodiments, inventive compounds exhibit IC₅₀ values lessthan 7.5 μM. In certain embodiments, inventive compounds exhibit IC₅₀values less than 5 μM. In certain other embodiments, inventive compoundsexhibit IC₅₀ values less than 2.5 μM. In certain embodiments, inventivecompounds exhibit IC₅₀ values less than 1 μM. In certain embodiments,inventive compounds exhibit IC₅₀ values less than 0.75 μM. In certainembodiments, inventive compounds exhibit IC₅₀ values less than 0.5 μM.In certain embodiments, inventive compounds exhibit IC₅₀ values lessthan 0.25 μM. In certain embodiments, inventive compounds exhibit IC₅₀values less than 0.1 μM. In certain other embodiments, inventivecompounds exhibit IC₅₀ values less than 75 nM. In certain otherembodiments, inventive compounds exhibit IC₅₀ values less than 50 nM. Incertain other embodiments, inventive compounds exhibit IC₅₀ values lessthan 25 nM. In certain other embodiments, inventive compounds exhibitIC₅₀ values less than 10 nM. In other embodiments, exemplary compoundsexhibit IC₅₀ values less than 7.5 nM. In other embodiments, exemplarycompounds exhibit IC₅₀ values less than 5 nM.

[0231] As discussed above, compounds of the invention exhibitimmunomodulatory activity and exhibit activity for the inhibition ofangiogenesis through inhibition of receptor tyrosine kinases. As such,the inventive compounds as useful for the treatment of a variety ofdisorders, including, but not limited to, sepsis, glomerulonephropathy,rheumatoid arthritis (including ankylosing spondylitis), psoriaticarthritis, osteoarthritis, osteoporosis, allergic rhinitis, ocularinflammation, inflammatory bowel disease, atopic dermatitis, psoriasis,asthma, Crohn's disease, ulcerative colitis, inflammatory pulmonarydisease, hepatitis, autoimmune disorders, diabetes, AIDS, solid tumorcancers, Leukemia, lymphomas, non-hodgkin's B-cell lymphomas, chronicallymphocytic leukemia (CLL), multiple myeloma, systemic lupuserythematosus, allograft rejection/graft versus host disease, eczema,uticaria, myasthenia gravis, idiopathic thrombocytopenia purpura,cardiovascular disease (e.g., myocardial infarction, atherosclerosis),hepatitis, productive nephritis, adenovirus, diseases/disorders of thecentral nervous system (stroke, Alzheimer's disease, epilepsy) and forthe treatment of the symptoms of malaria, to name a few. In certainembodiments, compounds of the invention are particularly useful for thetreatment of rheumatoid arthritis, psoriasis, multiple sclerosis, asthmaand cancer.

[0232] Rheumatoid Arthritis is a chronic syndrome characterized bynonspecific, usually symmetric inflammation of the peripheral joints,potentially resulting in progressive destruction of articular andperiarticular structures, with or without generalized manifestations(See, generally, The Merck Manual, 1999, Seventeenth Ed. Merck & Co.,the entire contents of which are hereby incorporated by reference).Studies in the past established that presence of inflammatory cells andpro-inflammatory cytokines, such as TNFα, IL-1β are abundant in thediseased synovium. Increased macrophage-derived lining cells areprominent along with some lymphocytes and vascular changes in earlydisease. Although there is not a cure, reduction of circulatorypro-inflammatory cytokines (e.g. TNFα, IL-1β) through intervention ofbiological agents, such as Enbrel, Remicade or Anakinra demonstratedefficacy in reduction of symptons and retarding the disease progressionin clinical trials. Thus developing of an agent such as described inthis patent in modulation of pro-inflammatory cytokines through NF-κBinhibition could bring great benefit to RA patients.

[0233] Psoriasis is a disorder for which there is no curative therapy,although in most cases acute attacks can be controlled. Psoriasis is achronic, recurrent disease characterized by dry, well-circumscribed,silvery, scaling papules and plaques of various sizes, and hastraditionally been attributed to increased epidermal cell proliferationand concomitant dermal inflammation. The response of psoriasis to theimmunosuppressive drug cyclosporine suggests that the primary pathogenicfactor may be immunologic. Proliferation of epidermal cells has beenalso linked to AP-1 activation via stimulation from injury, radiation orstress to the skin (see, P. Angel et al., “Function and regulation ofAP-1 subunits in skin physiology and pathology”, Oncogene, 2001,20:2413-2423; and A. Grandjean-Laquerriere et al., “Relativecontribution of NF-kB and AP-1 in the modulation by Curcumin andpyrrolidine dithiocarbamate of the UVB-induced cytokine expression bykeratinocytes”, Cytokine, 2002, 18(3): 168-177, each of which is herebyincorporated by reference in its entirety). Currently availabletreatment regimens for psoriasis include the use of lubricants,keratolytics, topical cortisosteroids, sunlight, topical vitamin Dderivatives, anthralin, and systemic antimetabolites (e.g.,methotrexate), immunosuppressive drugs (e.g., cyclosporine, tacrolimus,mycophenolate, and mofetil). However, immunosuppressive drugs are notyet approved for the treatment of psoriasis and other drugs, includingcorticosteriods, have severe side effects, including exacerbations orpustular lesions (See, generally, The Merck Manual, 1999, SeventeenthEd. Merck & Co., the entire contents of which are hereby incorporated byreference). This invention is certainly applicable to this disease aswell as a host of other related diseases, such as, psoriatic arthritis,ankylosing spondylitis, just to name a few.

[0234] Asthma is also believed to involve immunologic abnormalities andincreased inflammatory responses. Similarly to psoriasis, there is nocurative therapy. Thus the development of novel therapies such as this,preferably safe and curative, is desirable. This is also applied torelated immunologic disorders such as, graft rejection, SLE etc.

[0235] Angiogenesis, or the formation of new blood vessels out ofpre-existing capillaries, is a sequence of events that is fundamental tomany physiologic and pathologic processes such as cancer, ischemicdiseases, and chronical inflammation. With the identification of severalproangiogenic molecules such as vascular endothelial cell growth factor(VEGF), the fibroblast growth factors (FGFs) (see, Angiogenesis:Potentials for Pharmacologic Intervention in the Treatment of Cancer,Cardiovascular Diseases, and chronic Inflammation, PharmacologicalReviews, 2000, 52, 253). Thus, inhibition of receptor tyrosine kinase(such as VEGFr) activity has been subjects of various ongoing clinicaltrails. Certain compounds in this invention showed potent VEGFrinhibition. Thus, such application is expected.

[0236] As discussed herein, compounds of the invention inhibit theproduction of various pro-inflammatory and/or immunologic cytokines suchas TNFα, IL-1, IL-6, IL-8, IL-2 etc, and also inhibit the production ofvarious pro-inflammatory molecules under the regulation of NF-κB pathwaysuch as prostaglandins produced from COX-2, ICAM-1 and MMP-1 and 3 etc.

[0237] Elevated levels of proinflammatory cytokines are implicated inmany disease states, including rheumatoid arthritis (Dinarello, C. A.,et al. 1984, Rev. Infect. Disease 6:51; Maini, R. E. 1999, The Lancet354:1932; Weinblatt, M. E. 1999, New Eng. J. Med. 340:253),osteoarthritis (Pelletier and Pelletier 1989, J. Rheum. 16:19;Pelletier, et al. 1993, Am. J. Path. 142:95; Farahat, et al. 1993, Ann.Rheum. Dis. 52:870; Tiku, et al. 1992, Cell Immunol. 140:1; Webb, et al.1997, O. & C. 5:427; Westacott, et al. 2000, O. & C. 8:213), diabetes(McDaniel, et al 1996, Proc. Soc. Exp. Biol. Med. 211:24), HIV/AIDS(Kreuzer, et al. 1997, Clin. Exp. Immunol. 45:559), acute and chronicinflammatory diseases, such as the inflammatory reaction induced byendotoxin or inflammatory bowel disease, Crohn's disease and ulcerativecolitis (Rankin, E. C. C., et al. 1997, British J. Rheum. 35:334; Stack,W. A., et al. 1997, The Lancet 349:521); congestive heart failure Han etal. 2000, Trends Cardiovasc. Med. 10:19; Hunter et al 1999, N. Engl. J.Med. 341:1276; Behr et al. 2000, Circ. 102:II-289; Shimamoto et al.2000, Circ:102:II-289; Aukrust et al. 1999, Am. J. Cardiol. 83:376,hypertension (Singh, et al. 1996 J. Hypertension 9:867), chronicobstructive pulmonary disease, septic shock syndrome (Dinarello, C. A.1995, Nutrition 11:492), tuberculosis, adult respiratory distress,asthma (Renzetti, et al. Inflammation Res. 46:S143), atherosclerosis(Elhage, et al. 1998, Circulation 97:242), muscle degeneration,periodontal disease (Howells 1995, Oral Dis. 1:266), cachexia, Reiter'ssyndrome, gout, acute synovitis, eating disorders including anorexia andbulimia nervosa (Holden, et al. 1996, Med Hypothesis 47:423), fever,malaise, myalgia and headaches (Beisel 1995 Am. J. Clin. Nutr. 62:813).Inhibition of proinflammatory cytokine production, therefore, may offerthe opportunity to treat or prevent a wide range of diseases andconditions involving elevated levels of proinflammatory cytokines.

[0238] Although psoriasis is a localized skin condition, it might becaused and/or facilitated by systemic proinflammatory cytokines.Accordingly, without wishing to be bound to any particular theory, it isproposed that the inventive compounds might be surprisingly moreeffective for the treatment of psoriasis if made systemically available.Thus, in another aspect, the invention provides a method of treatingpsoriasis comprising systemically administering to a subject in needthereof an effective amount of a compound of the invention, optionallywith a pharmaceutically acceptable carrier. As used herein, “systemicadministration” refers to any means by which a compound of the inventioncan be made systemically available. For example, systemic administrationencompasses enteral (e.g., oral and rectal) and parenteral methods ofadministration. In certain embodiments, systemic administrationencompasses intravenous administration, intraperitoneal administration,intramuscular administration, intracoronary administration,intraarterial administration (e.g., into a carotid artery), intradermaladministration, subcutaneous administration, transdermal delivery,intratracheal administration, subcutaneous administration,intraarticular administration, intraventricular administration,inhalation (e.g., aerosol), intracerebral, nasal, naval, oral,intraocular, pulmonary administration, impregnation of a catheter, bysuppository and direct injection into a tissue, or systemically absorbedtopical or mucosal administration. Mucosal administration includesadministration to the respiratory tissue, e.g., by inhalation, nasaldrops, ocular drop, etc.; anal or vaginal routes of administration,e.g., by suppositories; and the like.

[0239] It will be appreciated that the inventive compound may beadministered systemically in dosage forms, formulations or e.g. suitabledelivery devices or implants containing conventional, non-toxicpharmaceutically acceptable carriers and adjuvants such that thecompound effectiveness is optimized. For example, the inventive compoundmay be formulated together with appropriate excipients into apharmaceutical composition, which, upon administration of thecomposition to the subject, systemically releases the active substancein a controlled manner. Alternatively, or additionally, compound dosageform designs may be optimized so as to increase the compoundeffectiveness upon administration. The above strategies (i.e., dosageform design and rate control of drug input), when used alone or incombination, can result in a significant increase in compoundeffectiveness and are considered part of the invention.

[0240] As discussed above, the inventive compounds also find use in theprevention of restenosis of blood vessels subject to traumas such asangioplasty and stenting. For example, it is contemplated that thecompounds of the invention will be useful as a coating for implantedmedical devices, such as tubings, shunts, catheters, artificialimplants, pins, electrical implants such as pacemakers, and especiallyfor arterial or venous stents, including balloon-expandable stents. Incertain embodiments inventive compounds may be bound to an implantablemedical device, or alternatively, may be passively adsorbed to thesurface of the implantable device. In certain other embodiments, theinventive compounds may be formulated to be contained within, or,adapted to release by a surgical or medical device or implant, such as,for example, stents, sutures, indwelling catheters, prosthesis, and thelike.

[0241] In certain exemplary embodiments, the inventive compounds may beused as coating for stents. A stent is typically an open tubularstructure that has a pattern (or patterns) of apertures extending fromthe outer surface of the stent to the lumen. It is commonplace to makestents of biocompatible metallic materials, with the patterns cut on thesurface with a laser machine. The stent can be electro-polished tominimize surface irregularities since these irregularities can triggeran adverse biological response. However, stents may still stimulateforeign body reactions that result in thrombosis or restenosis. To avoidthese complications, a variety of stent coatings and compositions havebeen proposed in the prior art literature both to reduce the incidenceof these complications or other complications and restore tissuefunction by itself or by delivering therapeutic compound to the lumen.For example, drugs having antiproliferative and anti-inflammatoryactivities have been evaluated as stent coatings, and have shown promisein preventing retenosis (See, for example, Presbitero P. et al., “Drugeluting stents do they make the difference?”, Minerva Cardioangiol,2002, 50(5):431-442; Ruygrok P. N. et al., “Rapamycin in cardiovascularmedicine”, Intern. Med. J., 2003, 33(3):103-109; and Marx S. O. et al.,“Bench to bedside: the development of rapamycin and its application tostent restenosis”, Circulation, 2001, 104(8):852-855, each of thesereferences is incorporated herein by reference in its entirety).Accordingly, without wishing to be bound to any particular theory,Applicant proposes that inventive compounds having anti-inflammatoryand/or antiproliferative effects can be used as stent coatings and/or instent drug delivery devices, inter alia for the prevention of restenosisor reduction of restenosis rate. A variety of compositions and methodsrelated to stent coating and/or local stent drug delivery for preventingrestenosis are known in the art (see, for example, U.S. Pat. Nos.6,517,889; 6,273,913; 6,258,121; 6,251,136; 6,248,127; 6,231,600;6,203,551; 6,153,252; 6,071,305; 5,891,507; 5,837,313 and published U.S.patent application No. US2001/0027340, each of which is incorporatedherein by reference in its entirety). For example, stents may be coatedwith polymer-drug conjugates by dipping the stent in polymer-drugsolution or spraying the stent with such a solution. In certainembodiment, suitable materials for the implantable device includebiocompatible and nontoxic materials, and may be chosen from the metalssuch as nickel-titanium alloys, steel, or biocompatible polymers,hydrogels, polyurethanes, polyethylenes, ethylenevinyl acetatecopolymers, etc. In certain embodiments, the inventive compound, iscoated onto a stent for insertion into an artery or vein followingballoon angioplasty.

[0242] The invention may be described therefore, in certain broadaspects as a method of inhibiting arterial restenosis or arterialocclusion following vascular trauma comprising administering to asubject in need thereof, a composition comprising an inventive compoundconjugated to a suitable polymer or polymeric material. In the practiceof the method, the subject may be a coronary bypass, vascular surgery,organ transplant or coronary or any other arterial angioplasty patient,for example, and the composition may be administered directly,intravenously, or even coated on a stent to be implanted at the sight ofvascular trauma.

[0243] In another aspect, the invention encompasses implants andsurgical or medical devices, including stents and grafts, coated with orotherwise constructed to contain and/or release any of the inventivecompounds disclosed herein. In certain embodiments, the compounds haveanti-inflammatory and/or antiproliferative activities. In certain otherembodiments, the compounds inhibit smooth muscle cell proliferation.Representative examples of the inventive implants and surgical ormedical devices include cardiovascular devices (e.g., implantable venouscatheters, venous ports, tunneled venous catheters, chronic infusionlines or ports, including hepatic artery infusion catheters, pacemakerwires, implantable defibrillators); neurologic/neurosurgical devices(e.g., ventricular peritoneal shunts, ventricular atrial shunts, nervestimulator devices, dural patches and implants to prevent epiduralfibrosis post-laminectomy, devices for continuous subarachnoidinfusions); gastrointestinal devices (e.g, chronic indwelling catheters,feeding tubes, portosystemic shunts, shunts for ascites, peritonealimplants for drug delivery, peritoneal dialysis catheters, implantablemeshes for hernias, suspensions or solid implants to prevent surgicaladhesions, including meshes); genitourinary devices (e.g., uterineimplants, including intrauterine devices (IUDs) and devices to preventendometrial hyperplasia, fallopian tubal implants, including reversiblesterilization devices, fallopian tubal stents, artificial sphincters andperiurethral implants for incontinence, ureteric stents, chronicindwelling catheters, bladder augmentations, or wraps or splints forvasovasostomy); phthalmologic implants (e.g., multino implants and otherimplants for neovascular glaucoma, drug eluting contact lenses forpterygiums, splints for failed dacrocystalrhinostomy, drug elutingcontact lenses for corneal neovascularity, implants for diabeticretinopathy, drug eluting contact lenses for high risk cornealtransplants); otolaryngology devices (e.g., ossicular implants,Eustachian tube splints or stents for glue ear or chronic otitis as analternative to transtempanic drains); plastic surgery implants (e.g.,prevention of fibrous contracture in response to gel- orsaline-containing breast implants in the subpectoral or subglandularapproaches or post-mastectomy, or chin implants), and orthopedicimplants (e.g., cemented orthopedic prostheses).

[0244] Implants and other surgical or medical devices may be coated with(or otherwise adapted to release) compositions of the present inventionin a variety of manners, including for example: (a) by directly affixingto the implant or device an inventive compound or composition (e.g., byeither spraying the implant or device with a polymer/drug film, or bydipping the implant or device into a polymer/drug solution, or by othercovalent or noncovalent means); (b) by coating the implant or devicewith a substance such as a hydrogel which will in turn absorb theinventive compound or composition; (c) by interweaving inventivecompound- or composition-coated thread (or the polymer itself formedinto a thread) into the implant or device; (d) by inserting the implantor device into a sleeve or mesh which is comprised of or coated with aninventive compound or composition; (e) constructing the implant ordevice itself with an inventive compound or composition; or (f) byotherwise adapting the implant or device to release the inventivecompound. In certain embodiments, the composition should firmly adhereto the implant or device during storage and at the time of insertion.The inventive compound or composition should also preferably not degradeduring storage, prior to insertion, or when warmed to body temperatureafter insertion inside the body (if this is required). In addition, itshould preferably coat the implant or device smoothly and evenly, with auniform distribution of inventive compound, while not changing the stentcontour. Within preferred embodiments of the invention, the inventiveimplant or device should provide a uniform, predictable, prolongedrelease of the inventive compound or composition into the tissuesurrounding the implant or device once it has been deployed. Forvascular stents, in addition to the above properties, the compositionshould not render the stent thrombogenic (causing blood clots to form),or cause significant turbulence in blood flow (more than the stentitself would be expected to cause if it was uncoated).

[0245] In the case of stents, a wide variety of stents may be developedto contain and/or release the inventive compounds or compositionsprovided herein, including esophageal stents, gastrointestinal stents,vascular stents, biliary stents, colonic stents, pancreatic stents,ureteric and urethral stents, lacrimal stents, Eustachian tube stents,fallopian tube stents and tracheal/bronchial stents (See, for example,U.S. Pat. No. 6,515,016, the entire contents of which are incorporatedherein by reference). Stents may be readily obtained from commercialsources, or constructed in accordance with well-known techniques.Representative examples of stents include those described in U.S. Pat.No. 4,768,523, entitled “Hydrogel Adhesive”; U.S. Pat. No. 4,776,337,entitled “Expandable Intraluminal Graft, and Method and Apparatus forImplanting and Expandable Intraluminal Graft”; U.S. Pat. No. 5,041,126entitled “Endovascular Stent and Delivery System”; U.S. Pat. No.5,052,998 entitled “Indwelling Stent and Method of Use”; U.S. Pat. No.5,064,435 entitled “Self-Expanding Prosthesis Having Stable AxialLength”; U.S. Pat. No. 5,089,606, entitled “Water-insolublePolysaccharide Hydrogel Foam for Medical Applications”; U.S. Pat. No.5,147,370, entitled “Nitinol Stent for Hollow Body Conduits”; U.S. Pat.No. 5,176,626, entitled “Indwelling Stent”; U.S. Pat. No. 5,213,580,entitled “Biodegradable Polymeric Endoluminal Sealing Process”; and U.S.Pat. No. 5,328,471, entitled “Method and Apparatus for Treatment ofFocal Disease in Hollow Tubular Organs and Other Tissue Lumens.”

[0246] As discussed above, the stent coated with (or otherwise adaptedto release) compositions of the present invention may be used toeliminate a vascular obstruction and prevent restenosis or reduce therate of restenosis. Within other aspects of the present invention,stents coated with (or otherwise adapted to release) compositions of thepresent invention are provided for expanding the lumen of a bodypassageway. Specifically, a stent having a generally tubular structure,and a surface coated with (or otherwise adapted to release) an inventivecompound or composition may be inserted into the passageway, such thatthe passageway is expanded. In certain embodiments, the stent coatedwith (or otherwise adapted to release) compositions of the presentinvention may be used to eliminate a biliary, gastrointestinal,esophageal, trachealibronchial, urethral or vascular obstruction.

[0247] In another aspect of the invention, methods for the treatment ofimmune disorders and cancer are provided comprising administering atherapeutically effective amount of a compound of formula (I), asdescribed herein, to a subject in need thereof. In certain embodiments,the inventive compounds are useful for the treatment of rheumatoidarthritis, psoriasis, multiple sclerosis, asthma and cancer. It will beappreciated that the compounds and compositions, according to the methodof the present invention, may be administered using any amount and anyroute of administration effective for the treatment of inflammatorydisorders, including but not limited to rheumatoid arthritis, psoriasis,multiple sclerosis, asthma and cancer. Thus, the expression “effectiveamount” as used herein, refers to a sufficient amount of agent toinhibit the growth of tumor cells, or refers to a sufficient amount toreduce the effects of rheumatoid arthritis, psoriasis, asthma andcancer, (or any inflammatory response or disorder). The exact amountrequired will vary from subject to subject, depending on the species,age, and general condition of the subject, the severity of the diseases,the particular anticancer agent, its mode of administration, and thelike. The compounds of the invention are preferably formulated in dosageunit form for ease of administration and uniformity of dosage. Theexpression “dosage unit form” as used herein refers to a physicallydiscrete unit of therapeutic agent appropriate for the patient to betreated. It will be understood, however, that the total daily usage ofthe compounds and compositions of the present invention will be decidedby the attending physician within the scope of sound medical judgment.The specific therapeutically effective dose level for any particularpatient or organism will depend upon a variety of factors including thedisorder being treated and the severity of the disorder; the activity ofthe specific compound employed; the specific composition employed; theage, body weight, general health, sex and diet of the patient; the timeof administration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed; andlike factors well known in the medical arts (see, for example, Goodmanand Gilman's, “The Pharmacological Basis of Therapeutics”, TenthEdition, A. Gilman, J. Hardman and L. Limbird, eds., McGraw-Hill Press,155-173, 2001, which is incorporated herein by reference in itsentirety).

[0248] In certain other embodiments, methods are provided for using theinventive implants and other surgical or medical devices coated with (orotherwise adapted to release) compounds and compositions of the presentinvention. In certain embodiments, methods are provided for preventingrestenosis, comprising inserting a stent into an obstructed bloodvessel, the stent having a generally tubular structure, the surface ofthe structure being coated with (or otherwise adapted to release) aninventive compound or composition, such that the obstruction iseliminated and the inventive compound or composition is delivered inamounts effective to prevent restenosis. In other embodiments, methodsare provided for preventing restenosis, comprising inserting a stentinto an obstructed blood vessel, the stent having a generally tubularstructure, the surface of the structure being coated with (or otherwiseadapted to release) an inventive compound or composition, such that theobstruction is eliminated and the inventive compound or composition isdelivered in amounts effective to inhibit smooth muscle cellproliferation.

[0249] Within other aspects of the present invention, methods areprovided for expanding the lumen of a body passageway, comprisinginserting a stent into the passageway, the stent having a generallytubular structure, the surface of the structure being coated with (orotherwise adapted to release) an inventive compound or composition, suchthat the passageway is expanded. In certain embodiments, the lumen of abody passageway is expanded in order to eliminate a biliary,gastrointestinal, esophageal, tracheal/bronchial, urethral and/orvascular obstruction.

[0250] In certain embodiments, methods are provided for eliminatingbiliary obstructions, comprising inserting a biliary stent into abiliary passageway, the stent having a generally tubular structure, thesurface of the structure being coated with (or otherwise adapted torelease) an inventive compound or composition, such that the biliaryobstruction is eliminated. Briefly, tumor overgrowth of the conimon bileduct results in progressive cholestatic jaundice which is incompatiblewith life. Generally, the biliary system which drains bile from theliver into the duodenum is most often obstructed by (1) a tumor composedof bile duct cells (cholangiocarcinoma), (2) a tumor which invades thebile duct (e.g., pancreatic carcinoma), or (3) a tumor which exertsextrinsic pressure and compresses the bile duct (e.g., enlarged lymphnodes). Both primary biliary tumors, as well as other tumors which causecompression of the biliary tree may be treated utilizing stents Implantsand other surgical or medical devices may be coated with (or otherwiseadapted to release) compositions of the present invention. One exampleof primary biliary tumors are adenocarcinomas (which are also calledKlatskin tumors when found at the bifurcation of the common hepaticduct). These tumors are also referred to as biliary carcinomas,choledocholangiocarcinomas, or adenocarcinomas of the biliary system.Benign tumors which affect the bile duct (e.g., adenoma of the biliarysystem), and, in rare cases, squamous cell carcinomas of the bile ductand adenocarcinomas of the gallbladder, may also cause compression ofthe biliary tree and therefore, result in biliary obstruction.Compression of the biliary tree is most commonly due to tumors of theliver and pancreas which compress and therefore obstruct the ducts. Mostof the tumors from the pancreas arise from cells of the pancreaticducts. This is a highly fatal form of cancer (5% of all cancer deaths;26,000 new cases per year in the U.S.) with an average of 6 monthssurvival and a 1 year survival rate of only 10%. When these tumors arelocated in the head of the pancreas they frequently cause biliaryobstruction, and this detracts significantly from the quality of life ofthe patient. While all types of pancreatic tumors are generally referredto as “carcinoma of the pancreas” there are histologic subtypesincluding: adenocarcinoma, adenosquamous carcinoma, cystadenocarcinoma,and acinar cell carcinoma. Hepatic tumors, as discussed above, may alsocause compression of the biliary tree, and therefore cause obstructionof the biliary ducts.

[0251] In certain embodiments, a biliary stent is first inserted into abiliary passageway in one of several ways: from the top end by insertinga needle through the abdominal wall and through the liver (apercutaneous transhepatic cholangiogram or “PTC”); from the bottom endby cannulating the bile duct through an endoscope inserted through themouth, stomach, and duodenum (an endoscopic retrograde cholangiogram or“ERCP”); or by direct incision during a surgical procedure. In certainembodiments, a preinsertion examination, PTC, ERCP, or directvisualization at the time of surgery is performed to determine theappropriate position for stent insertion. A guidewire is then advancedthrough the lesion, and over this a delivery catheter is passed to allowthe stent to be inserted in its collapsed form. If the diagnostic examwas a PTC, the guidewire and delivery catheter is inserted via theabdominal wall, while if the original exam was an ERCP the stent may beplaced via the mouth. The stent is then positioned under radiologic,endoscopic, or direct visual control taking particular care to place itprecisely across the narrowing in the bile duct. The delivery catheteris then removed leaving the stent standing as a scaffolding which holdsthe bile duct open. A further cholangiogram may be performed to documentthat the stent is appropriately positioned.

[0252] In certain embodiments, methods are provided for eliminatingesophageal obstructions, comprising inserting an esophageal stent intoan esophagus, the stent having a generally tubular structure, thesurface of the structure being coated with (or otherwise adapted torelease) an inventive compound or composition, such that the esophagealobstruction is eliminated. Briefly, the esophagus is the hollow tubewhich transports food and liquids from the mouth to the stomach. Cancerof the esophagus or invasion by cancer arising in adjacent organs (e.g.,cancer of the stomach or lung) results in the inability to swallow foodor saliva. In certain embodiments, a preinsertion examination, usually abarium swallow or endoscopy is performed in order to determine theappropriate position for stent insertion. A catheter or endoscope maythen be positioned through the mouth, and a guidewire is advancedthrough the blockage. A stent delivery catheter is passed over theguidewire under radiologic or endoscopic control, and a stent is placedprecisely across the narrowing in the esophagus. A post-insertionexamination, usually a barium swallow x-ray, may be utilized to confirmappropriate positioning.

[0253] In certain embodiments, methods are provided for eliminatingcolonic obstructions, comprising inserting a colonic stent into a colon,the stent having a generally tubular structure, the surface of thestructure being coated with (or otherwise adapted to release) aninventive compound or composition, such that the colonic obstruction iseliminated. Briefly, the colon is the hollow tube which transportsdigested food and waste materials from the small intestines to the anus.Cancer of the rectum and/or colon or invasion by cancer arising inadjacent organs (e.g., cancer of the uterus, ovary, bladder) results inthe inability to eliminate feces from the bowel. In certain embodiments,a preinsertion examination, usually a barium enema or colonoscopy isperformed in order to determine the appropriate position for stentinsertion. A catheter or endoscope may then be positioned through theanus, and a guidewire is advanced through the blockage. A stent deliverycatheter is passed over the guidewire under radiologic or endoscopiccontrol, and a stent is placed precisely across the narrowing in thecolon or rectum. A post-insertion examination, usually a barium enemax-ray, may be utilized to confirm appropriate positioning.

[0254] In certain embodiments, methods are provided for eliminatingtracheal/bronchial obstructions, comprising inserting atracheal/bronchial stent into a trachea or bronchi, the stent having agenerally tubular structure, the surface of the structure being coatedwith (or otherwise adapted to release) an inventive compound orcomposition, such that the tracheal/bronchial obstruction is eliminated.Briefly, the trachea and bronchi are tubes which carry air from themouth and nose to the lungs. Blockage of the trachea by cancer, invasionby cancer arising in adjacent organs (e.g., cancer of the lung), orcollapse of the trachea or bronchi due to chondromalacia (weakening ofthe cartilage rings) results in inability to breathe. In certainembodiments, preinsertion examination, usually an endoscopy, isperformed in order to determine the appropriate position for stentinsertion. A catheter or endoscope is then positioned through the mouth,and a guidewire advanced through the blockage. A delivery catheter isthen passed over the guidewire in order to allow a collapsed stent to beinserted. The stent is placed under radiologic or endoscopic control inorder to place it precisely across the narrowing. The delivery cathetermay then be removed leaving the stent standing as a scaffold on its own.A post-insertion examination, usually a bronchoscopy may be utilized toconfirm appropriate positioning.

[0255] In certain embodiments, methods are provided for eliminatingurethral obstructions, comprising inserting a urethral stent into aurethra, the stent having a generally tubular structure, the surface ofthe structure being coated with (or otherwise adapted to release) aninventive compound or composition, such that the urethral obstruction iseliminated. Briefly, the urethra is the tube which drains the bladderthrough the penis. Extrinsic narrowing of the urethra as it passesthrough the prostate, due to hypertrophy of the prostate, occurs invirtually every man over the age of 60 and causes progressive difficultywith urination. In certain embodiments, a preinsertion examination,usually an endoscopy or urethrogram is first performed in order todetermine the appropriate position for stent insertion, which is abovethe external urinary sphincter at the lower end, and close to flush withthe bladder neck at the upper end. An endoscope or catheter is thenpositioned through the penile opening and a guidewire advanced into thebladder. A delivery catheter is then passed over the guidewire in orderto allow stent insertion. The delivery catheter is then removed, and thestent expanded into place. A post-insertion examination, usuallyendoscopy or retrograde urethrogram, may be utilized to confirmappropriate position.

[0256] In certain embodiments, methods are provided for eliminatingvascular obstructions, comprising inserting a vascular stent into ablood vessel, the stent having a generally tubular structure, thesurface of the structure being coated with (or otherwise adapted torelease) an inventive compound or composition, such that the vascularobstruction is eliminated. Briefly, stents may be placed in a wide arrayof blood vessels, both arteries and veins, to prevent recurrent stenosisat the site of failed angioplasties, to treat narrowings that wouldlikely fail if treated with angioplasty, and to treat post-surgicalnarrowings (e.g., dialysis graft stenosis). Suitable sites include, butare not limited to, the iliac, renal, and coronary arteries, thesuperior vena cava, and in dialysis grafts. In certain embodiments,angiography is first performed in order to localize the site forplacement of the stent. This is typically accomplished by injectingradiopaque contrast through a catheter inserted into an artery or veinas an x-ray is taken. A catheter may then be inserted eitherpercutaneously or by surgery into the femoral artery, brachial artery,femoral vein, or brachial vein, and advanced into the appropriate bloodvessel by steering it through the vascular system under fluoroscopicguidance. A stent may then be positioned across the vascular stenosis. Apost-insertion angiogram may also be utilized in order to confirmappropriate positioning.

[0257] In certain other embodiments, compounds of the invention areuseful for reducing photodamage, and thus, the invention furtherprovides a method for treating photoaging-related disorders/conditions.Photoaging is a term used to describe the changes in appearance andfunction of skin as a result of repeated exposure to sunlight. Theultraviolet (UV) component of sunlight, particularly middle UV (calledUVB, 290-320 nm wavelength) is the principal causative agent ofphotoaging. The extent of UVB exposure required to cause photoaging isnot currently known. Repeated exposure to UVB at levels that causeerythema and tanning are, however, commonly associated with photoaging.Clinically, photoaging is characterized by coarseness, wrinkling,mottled pigmentation, sallowness, laxity, telangiectasia, lentigines,purpura and easy bruising, atrophy, fibrotic depigmented areas, andultimately premalignant and malignant neoplasms. Photoaging commonlyoccurs in skin that is habitually exposed to sunlight such as the face,ears, bald areas of the scalp, neck, and hands.

[0258] Procedures for preventing photoaging of unaged skin and treatingalready photoaged skin are available. Sunscreens are commonly used toprevent photoaging of skin areas that are habitually exposed tosunlight. Sunscreens are topical preparations that absorb, reflect orscatter UV. Some are based on opaque particulate materials such as zincoxide, titanium oxide, clays and ferric chloride. Because suchpreparations are visible and occlusive many people consider these opaqueformulations cosmetically unacceptable. Other sunscreens containchemicals such a p-aminobenzoic acid (PABA), oxybenzone, dioxybenzone,ethylhexyl-methoxy cinnamide and butylmethoxydibenzoylmethane that arenonopaque and colorless because they do not absorb light of visiblewavelengths. While these nonopaque sunscreens may be more acceptablecosmetically they are still relatively short-lived and susceptible tobeing removed by washing or perspiration. Additionally all sunscreensreduce vitamin D production.

[0259] It is known that transcription factors AP-1 and NF-κB areactivated in mammalian cells exposed to UV light. It has also been shownthat inhibition of MAP kinase/ERK kinase 1 (MEK-1) significantlyinhibited UVB induced ERK activation (See, Chen et al., “Activation ofp38 MAP kinase and ERK are required for ultraviolet-B induced c-fos geneexpression in human keratinocytes”, Oncogene, 18:7469-7476, 1999; theentire contents of which are incorporated herein by reference).Accordingly, without wishing to be bound to any particular theory,Applicant proposes that the compounds of the invention may find use inthe treatment of skin damages caused by UVB exposure. For additionalreferences on MAP kinases and photoaging, see Li et al., “Rays andarrays: the transcriptional program in the response of human epidermalkeratotinocutes to UVB illumination”, The FASEB Journal express article10.1096/fj.01-01172fje, published online Sep. 17, 2001; U.S. Pat. No.5,837,224 and U.S. patent application Ser. No. 20020106339, each ofwhich is hereby incorporated by reference in its entirety.

[0260] Thus the invention provides compositions for preventing ortreating UVB-induced photodamage comprising an inventive compound; and apharmaceutically acceptable carrier. In certain embodiments, theinventive compound is present in an amount effective to inhibit Map/Erkkinase. In certain other embodiments, the inventive compositions furthercomprise a cosmetic ingredient. In certain exemplary embodiments, thecosmetic ingredient is a fragrance. In certain other exemplaryembodiments, the cosmetic ingredient is a sunscreen. In certainembodiments, the inventive compositions exist as pharmaceuticallyacceptable topical formulations.

[0261] The present invention additionally encompasses methods ofproviding protection against long-term UVB induced photodamage to asubject, said method comprising: administering to the subject in needthereof a composition comprising an inventive compound; and apharmaceutically acceptable carrier or diluent. In certain embodiments,the composition is administered topically. The present inventionadditionally encompasses methods of providing protection againstlong-term UVB induced photodamage to a subject, said method comprising:providing the subject with a composition comprising an inventivecompound; and providing the subject with instructions for using saidcomposition to prevent photodamage. In certain embodiments, thecomposition is formulated so that it may be administered topically. Incertain embodiments, the inventive compound is present in an amounteffective to inhibit Map/Erk kinase. In certain embodiments, theinstructions comprise directions to apply the composition to the skinprior to sun exposure. In certain exemplary embodiments, the compositionfurther comprises a cosmetic ingredient. In certain exemplaryembodiments, the cosmetic ingredient is a fragrance. In certain otherexemplary embodiments, the cosmetic ingredient is a sunscreen. Incertain embodiment, a method is provided for treating and/or preventingskin coarseness, wrinkling, mottled pigmentation, sallowness, laxity,telangiectasia, lentigines, purpura and easy bruising, atrophy, fibroticdepigmented areas, and ultimately premalignant and malignant neoplasms.In certain exemplary embodiments, the present invention provides amethod for treating and/or preventing wrinkles and/or skin cancer.

[0262] In certain embodiments, the present invention provides kits forpreventing long-term UVB induced photodamage in a subject, said kitcomprising: a composition comprising an inventive compound; andinstructions for using the composition to prevent photodamage. Incertain embodiments, the composition is formulated for topicaladministration. In certain embodiments, the inventive compound ispresent in an amount effective to inhibit Map/Erk kinase. In certainembodiments, the instructions comprise directions to apply thecomposition to the skin prior to sun exposure. In certain exemplaryembodiments, the composition further comprises a cosmetic ingredient. Incertain exemplary embodiments, the cosmetic ingredient is a fragrance.In certain other exemplary embodiments, the cosmetic ingredient is asunscreen.

[0263] Furthermore, after formulation with an appropriatepharmaceutically acceptable carrier or diluent in a desired dosage, thepharmaceutical compositions of this invention can be administered tohumans and other animals orally, rectally, parenterally,intracisternally, intravaginally, intraperitoneally, topically (as bypowders, ointments, creams or drops), bucally, as an oral or nasalspray, or the like, depending on the severity of the infection beingtreated. In certain embodiments, the compounds of the invention may beadministered at dosage levels of about 0.001 mg/kg to about 50 mg/kg,from about 0.01 mg/kg to about 25 mg/kg, or from about 0.1 mg/kg toabout 10 mg/kg of subject body weight per day, one or more times a day,to obtain the desired therapeutic effect. It will also be appreciatedthat dosages smaller than 0.001 mg/kg or greater than 50 mg/kg (forexample 50-100 mg/kg) can be administered to a subject. In certainembodiments, compounds are administered orally or parenterally.

[0264] Liquid dosage forms for oral administration include, but are notlimited to, pharmaceutically acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. In addition to the activecompounds, the liquid dosage forms may contain inert diluents commonlyused in the art such as, for example, water or other solvents,solubilizing agents and emulsifiers such as ethyl alcohol, isopropylalcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils(in particular, cottonseed, groundnut, corn, germ, olive, castor, andsesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycolsand fatty acid esters of sorbitan, and mixtures thereof. Besides inertdiluents, the oral compositions can also include adjuvants such aswetting agents, emulsifying and suspending agents, sweetening,flavoring, and perfuming agents.

[0265] Injectable preparations, for example, sterile injectable aqueousor oleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

[0266] The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

[0267] In order to prolong the effect of a drug, it is often desirableto slow the absorption of the drug from subcutaneous or intramuscularinjection. This may be accomplished by the use of a liquid suspension orcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionthat, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude (poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions, which are compatible with body tissues.

[0268] Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

[0269] Solid dosage forms for oral administration include capsules,tablets, pills, powders, and granules. In such solid dosage forms, theactive compound is mixed with at least one inert, pharmaceuticallyacceptable excipient or carrier such as sodium citrate or dicalciumphosphate and/or a) fillers or extenders such as starches, lactose,sucrose, glucose, mannitol, and silicic acid, b) binders such as, forexample, carboxymethylcellulose, alginates, gelatin,polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such asglycerol, d) disintegrating agents such as agar--agar, calciumcarbonate, potato or tapioca starch, alginic acid, certain silicates,and sodium carbonate, e) solution retarding agents such as paraffin, f)absorption accelerators such as quaternary ammonium compounds, g)wetting agents such as, for example, cetyl alcohol and glycerolmonostearate, h) absorbents such as kaolin and bentonite clay, and i)lubricants such as talc, calcium stearate, magnesium stearate, solidpolyethylene glycols, sodium lauryl sulfate, and mixtures thereof. Inthe case of capsules, tablets and pills, the dosage form may alsocomprise buffering agents.

[0270] Solid compositions of a similar type may also be employed asfillers in soft and hard-filled gelatin capsules using such excipientsas lactose or milk sugar as well as high molecular weight polyethyleneglycols and the like. The solid dosage forms of tablets, dragees,capsules, pills, and granules can be prepared with coatings and shellssuch as enteric coatings and other coatings well known in thepharmaceutical formulating art. They may optionally contain opacifyingagents and can also be of a composition that they release the activeingredient(s) only, or preferentially, in a certain part of theintestinal tract, optionally, in a delayed manner. Examples of embeddingcompositions that can be used include polymeric substances and waxes.Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polethylene glycols andthe like.

[0271] The active compounds can also be in microencapsulated form withone or more excipients as noted above. The solid dosage forms oftablets, dragees, capsules, pills, and granules can be prepared withcoatings and shells such as enteric coatings, release controllingcoatings and other coatings well known in the pharmaceutical formulatingart. In such solid dosage forms the active compound may be admixed withat least one inert diluent such as sucrose, lactose and starch. Suchdosage forms may also comprise, as in normal practice, additionalsubstances other than inert diluents, e.g., tableting lubricants andother tableting aids such as magnesium stearate and microcrystallinecellulose. In the case of capsules, tablets and pills, the dosage formsmay also comprise buffering agents. They may optionally containopacifying agents and can also be of a composition that they release theactive ingredient(s) only, or preferentially, in a certain part of theintestinal tract, optionally, in a delayed manner. Examples of embeddingcompositions, which can be used, include polymeric substances and waxes.

[0272] The present invention encompasses pharmaceutically acceptabletopical formulations of inventive compounds. The term “pharmaceuticallyacceptable topical formulation”, as used herein, means any formulationwhich is pharmaceutically acceptable for intradermal administration of acompound of the invention by application of the formulation to theepidermis. In certain embodiments of the invention, the topicalformulation comprises a carrier system. Pharmaceutically effectivecarriers include, but are not limited to, solvents (e.g., alcohols, polyalcohols, water), creams, lotions, ointments, oils, plasters, liposomes,powders, emulsions, microemulsions, and buffered solutions (e.g.,hypotonic or buffered saline) or any other carrier known in the art fortopically administering pharmaceuticals. A more complete listing ofart-known carriers is provided by reference texts that are standard inthe art, for example, Remington's Pharmaceutical Sciences, 16th Edition,1980 and 17th Edition, 1985, both published by Mack Publishing Company,Easton, Pa., the disclosures of which are incorporated herein byreference in their entireties. In certain other embodiments, the topicalformulations of the invention may comprise excipients. Anypharmaceutically acceptable excipient known in the art may be used toprepare the inventive pharmaceutically acceptable topical formulations.Examples of excipients that can be included in the topical formulationsof the invention include, but are not limited to, preservatives,antioxidants, moisturizers, emollients, buffering agents, solubilizingagents, other penetration agents, skin protectants, surfactants, andpropellants, and/or additional therapeutic agents used in combination tothe inventive compound. Suitable preservatives include, but are notlimited to, alcohols, quaternary amines, organic acids, parabens, andphenols. Suitable antioxidants include, but are not limited to, ascorbicacid and its esters, sodium bisulfite, butylated hydroxytoluene,butylated hydroxyanisole, tocopherols, and chelating agents like EDTAand citric acid. Suitable moisturizers include, but are not limited to,glycerine, sorbitol, polyethylene glycols, urea, and propylene glycol.Suitable buffering agents for use with the invention include, but arenot limited to, citric, hydrochloric, and lactic acid buffers. Suitablesolubilizing agents include, but are not limited to, quaternary ammoniumchlorides, cyclodextrins, benzyl benzoate, lecithin, and polysorbates.Suitable skin protectants that can be used in the topical formulationsof the invention include, but are not limited to, vitamin E oil,allatoin, dimethicone, glycerin, petrolatum, and zinc oxide.

[0273] In certain embodiments, the pharmaceutically acceptable topicalformulations of the invention comprise at least a compound of theinvention and a penetration enhancing agent. The choice of topicalformulation will depend or several factors, including the condition tobe treated, the physicochemical characteristics of the inventivecompound and other excipients present, their stability in theformulation, available manufacturing equipment, and costs constraints.As used herein the term “penetration enhancing agent” means an agentcapable of transporting a pharmacologically active compound through thestratum comeum and into the epidermis or dermis, preferably, with littleor no systemic absorption. A wide variety of compounds have beenevaluated as to their effectiveness in enhancing the rate of penetrationof drugs through the skin. See, for example, Percutaneous PenetrationEnhancers, Maibach H. I. and Smith H. E. (eds.), CRC Press, Inc., BocaRaton, Fla. (1995), which surveys the use and testing of various skinpenetration enhancers, and Buyuktimkin et al., Chemical Means ofTransdermal Drug Permeation Enhancement in Transdermal and Topical DrugDelivery Systems, Gosh T. K., Pfister W. R., Yum S. I. (Eds.),Interpharm Press Inc., Buffalo Grove, Ill. (1997). In certain exemplaryembodiments, penetration agents for use with the invention include, butare not limited to, triglycerides (e.g., soybean oil), aloe compositions(e.g., aloe-vera gel), ethyl alcohol, isopropyl alcohol,octolyphenylpolyethylene glycol, oleic acid, polyethylene glycol 400,propylene glycol, N-decylmethylsulfoxide, fatty acid esters (e.g.,isopropyl myristate, methyl laurate, glycerol monooleate, and propyleneglycol monooleate) and N-methyl pyrrolidone.

[0274] In certain embodiments, the compositions may be in the form ofointments, pastes, creams, lotions, gels, powders, solutions, sprays,inhalants or patches. In certain exemplary embodiments, formulations ofthe compositions according to the invention are creams, which mayfurther contain saturated or unsaturated fatty acids such as stearicacid, palmitic acid, oleic acid, palmito-oleic acid, cetyl or oleylalcohols, stearic acid being particularly preferred. Creams of theinvention may also contain a non-ionic surfactant, for example,polyoxy-40-stearate. In certain embodiments, the active component isadmixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, eardrops, and eye drops are also contemplated asbeing within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms are made by dissolving or dispensing thecompound in the proper medium. As discussed above, penetration enhancingagents can also be used to increase the flux of the compound across theskin. The rate can be controlled by either providing a rate controllingmembrane or by dispersing the compound in a polymer matrix or gel.

[0275] In certain embodiments, after application of the topicalformulation to the epidermis, the area may be covered with a dressing.The term “dressing”, as used herein, means a covering designed toprotect a topically applied drug formulation. “Dressing” includescoverings such as a bandage, which may be porous or non-porous andvarious inert coverings, e.g., a plastic film wrap or othernon-absorbent film. The term “dressing” also encompasses non-woven orwoven coverings, particularly elastomeric coverings, which allow forheat and vapor transport. These dressings allow for cooling of thetreated area, which provides for greater comfort.

[0276] In certain exemplary embodiments, pharmaceutically acceptabletopical formulations of the invention are contained in a patch that isapplied adjacent to the area of skin to be treated. As used herein a“patch” comprises at least a topical formulation and a covering layer,such that, the patch can be placed over the area of skin to be treated.Preferably, but not necessarily, the patch is designed to maximize drugdelivery through the stratum corneum and into the epidermis or dermis,reduce lag time, promote uniform absorption, and/or reduce mechanicalrub-off. In certain embodiments, when the intended use comprises thetreatment of a skin condition (e.g., psoriasis), the patch is designedto minimize absorption into the circulatory system. Preferably, thepatch components resemble the viscoelastic properties of the skin andconform to the skin during movement to prevent undue shear anddelamination. Advantages of a patch comprising the topical formulationof the invention over conventional methods of administration include (i)that the dose is controlled by the patch's surface area, (ii) constantrate of administration, (iii) longer duration of action (the ability ofto adhere to the skin for 1, 3, 7 days or longer), (iv) improved patientcompliance, (v) non-invasive dosing, and (vi) reversible action (i.e.,the patch can simply be removed).

[0277] In certain embodiments, a patch suitable for use with theinvention contains at least: (1) a backing layer and (2) a carrierformulated with a compound of the invention. Examples of patch systemssuitable for practicing the invention include, but are not limited to,matrix-type patches; reservoir-type patches; multi-laminatedrug-in-adhesive-type patches; and monolithic drug-in-adhesivetype-patch. See, for example Ghosh, T. K.; Pfister, W. R.; Yum, S. I.Transdermal and Topical Drug Delivery Systems, Interpharm Press, Inc. p.249-297, which is incorporated herein by reference in its entirety.These patches are well known in the art and generally availablecommercially.

[0278] The matrix patch comprises matrix containing an inventivecompound, an adhesive backing film overlay, and preferably, but notnecessarily, a release liner. In some cases, it may be necessary toinclude a impermeable layer to minimize drug migration into the backingfilm (e.g., U.S. Pat. No. 4,336,243, incorporated herein by reference).In certain embodiments, the matrix containing the inventive compound isheld against the skin by the adhesive overlay. Examples of suitablematrix materials include but are not limited to lipophilic polymers,such as polyvinyl chloride, polydimethylsiloxane, and hydrophilicpolymers like polyvinylpyrrolidone, polyvinyl alcohol, hydrogels basedon gelatin, or polyvinylpyrrolidone/polyethylene oxide mixtures.Suitable release liners include but are not limited to occlusive,opaque, or clear polyester films with a thin coating of pressuresensitive release liner (e.g., silicone-fluorsilicone, andperfluorcarbon based polymers.

[0279] The reservoir type patch design is characterized by a backingfilm coated with an adhesive, and a reservoir compartment comprising adrug formulation preferably, in the form of a solution or suspension,that is separated from the skin by a semipermeable membrane (e.g., U.S.Pat. No. 4,615,699, incorporated herein by reference). The adhesivecoated backing layer extends around the reservoir's boundaries toprovide a concentric seal with the skin and hold the reservoir adjacentto the skin.

[0280] The monolithic drug-in-adhesive patch design is characterized bythe inclusion of the drug formulation in the skin contacting adhesivelayer, a backing film and preferably, a release liner. The adhesivefunctions both to release the compound and adhere the compound matrix tothe skin. The drug-in-adhesive system does not require an adhesiveoverlay and thus the patch size is minimized. Also, drug-in-adhesivetype patches are thin and comfortable (e.g., U.S. Pat. No. 4,751,087,incorporated herein by reference).

[0281] The multi-laminate drug-in-adhesive patch design furtherincorporates an additional semi-permeable membrane between two distinctdrug-in-adhesive layers or multiple drug-in-adhesive layers under asingle backing film (Peterson, T. A. and Dreyer, S. J. Proceed. Intern.Symp. Control. Rel. Bioact. Mater. 21: 477-478, incorporated herein byreference).

[0282] Semi permeable membranes, useful with the reservoir ormulti-laminate patch, include thin non-porous ethylene vinyl acetatefilms or thin microporous films of polyethylene employed inmicrolaminate solid state reservoir patches.

[0283] Adhesives for use with the drug-in-adhesive type patches are wellknown in the art and a pratitioner skilled in the relevant art wouldknow how to select an adhesive suitable for the intended use. Examplesof adhesives include, but are not limited to, polyisobutylenes,silicones, and acrylics. Preferably, adhesives can function under a widerange of conditions, such as, high and low humidity, bathing, sweatingetc. Preferably the adhesive is a composition based on natural orsynthetic rubber; a polyacrylate such as, polybutylacrylate,polymethylacrylate, poly-2-ethylhexyl acrylate; polyvinylacetate;polydimethylsiloxane; pressure sensitive acrylic adhesives, for exampleDurotak.RTM. adhesives (e.g., Durotak.RTM. 2052, National Starch andChemicals) or hydrogels (e.g., high molecular weightpolyvinylpyrrolidone and oligomeric polyethylene oxide). The adhesivemay contain a thickener, such as a silica thickener (e.g., Aerosil,Degussa, Ridgefield Park, N.J.) or a crosslinker such as,aluminumacetylacetonate.

[0284] Backing films may be occlusive or permeable and may be derivedfrom synthetic polymers like polyolefin oils polyester, polyethylene,polyvinylidine chloride, and polyurethane or from natural materials likecotton, wool, etc. Occlusive backing films, such as syntheticpolyesters, result in hydration of the outer layers of the stratumcorneum while non-occlusive backings allow the area to breath (i.e.,promote water vapor transmission from the skin surface).

[0285] Selection of the appropriate dosage for the application site isan important consideration. The rate of compound intradermaladministration from the topical formulation or patch is a function ofskin permeability, and skin permeability has been shown to vary betweenanatomical sites depending on the thickness of the stratum comeum. Forexample, the permeability, in general, increases in order from planterfoot arch, lateral ankle, palm, ventral forearm, dorsal forearm, back,chest, thigh, abdomen, scalp, axilla, forehead, and scrotum (Wester, R.C. and Maibach, H. I. (1989) Regional variation in PercutaneousAbsorption: In Percutaneous Absorption, Mechanism, Methodology, DrugDelivery, 2^(nd) ed., Eds. R. L. Bronaugh and H. I. Maibach, MarcelDekker, Inc., New York, pp. 111-119 (incorporated herein by reference)).Typically, the dosages and dosing frequency will be determined by atrained medical professional.

[0286] It will also be appreciated that the compounds and pharmaceuticalcompositions of the present invention can be formulated and employed incombination therapies, that is, the compounds and pharmaceuticalcompositions can be formulated with or administered concurrently with,prior to, or subsequent to, one or more other desired therapeutics ormedical procedures. The particular combination of therapies(therapeutics or procedures) to employ in a combination regimen willtake into account compatibility of the desired therapeutics and/orprocedures and the desired therapeutic effect to be achieved. It willalso be appreciated that the therapies employed may achieve a desiredeffect for the same disorder (for example, an inventive compound may beadministered concurrently with another immunomodulatory agent,anticancer agent or agent useful for the treatment of psoriasis), orthey may achieve different effects (e.g., control of any adverseeffects).

[0287] For example, other therapies or anticancer agents that may beused in combination with the inventive compounds of the presentinvention include surgery, radiotherapy (in but a few examples,γ-radiation, neutron beam radiotherapy, electron beam radiotherapy,proton therapy, brachytherapy, and systemic radioactive isotopes, toname a few), endocrine therapy, biologic response modifiers(interferons, interleukins, and tumor necrosis factor (TNF) to name afew), hyperthermia and cryotherapy, agents to attenuate any adverseeffects (e.g., antiemetics), and other approved chemotherapeutic drugs,including, but not limited to, alkylating drugs (mechlorethamine,chlorambucil, Cyclophosphamide, Melphalan, Ifosfamide), antimetabolites(Methotrexate), purine antagonists and pyrimidine antagonists(6-Mercaptopurine, 5-Fluorouracil, Cytarabile, Gemcitabine), spindlepoisons (Vinblastine, Vincristine, Vinorelbine, Paclitaxel),podophyllotoxins (Etoposide, Irinotecan, Topotecan), antibiotics(Doxorubicin, Bleomycin, Mitomycin), nitrosoureas (Carmustine,Lomustine), inorganic ions (Cisplatin, Carboplatin), enzymes(Asparaginase), and hormones (Tamoxifen, Leuprolide, Flutamide, andMegestrol), to name a few. For a more comprehensive discussion ofupdated cancer therapies see, The Merck Manual, Seventeenth Ed. 1999,the entire contents of which are hereby incorporated by reference. Seealso the National Cancer Institute (CNI) website (www.nci.nih.gov) andthe Food and Drug Administration (FDA) website for a list of the FDAapproved oncology drugs (www.fda.gov/cder/cancer/druglistframe—SeeAppendix A).

[0288] In certain embodiments, the pharmaceutical compositions of thepresent invention further comprise one or more additionaltherapeutically active ingredients (e.g., chemotherapeutic and/orpalliative). For purposes of the invention, the term “Palliative” refersto treatment that is focused on the relief of symptoms of a diseaseand/or side effects of a therapeutic regimen, but is not curative. Forexample, palliative treatment encompasses painkillers, antinauseamedications and anti-sickness drugs. In addition, chemotherapy,radiotherapy and surgery can all be used palliatively (that is, toreduce symptoms without going for cure; e.g., for shrinking tumors andreducing pressure, bleeding, pain and other symptoms of cancer).

[0289] In certain embodiments, compounds of the invention are useful forthe treatment of psoriasis and pharmaceutical compositions containingthem may be administered in combination with any of the antipsoriatictherapies or therapeutic agents known in the art. For example, therapiesor antipsoriatic agents that may be used in combination with theinventive compounds of the present invention include Ultraviolet lighttreatment (e.g., sunlight), lubricants, keratolytics, emollients (e.g.,Aqueous Cream, E45, and Emulsifying ointment), ammoniated mercury,topical vitamin D analogs (e.g., Calcipotriol (Dovonex), Tacalcitol(Curatoderm)), dithranol (e.g., Dithrocream and Miconal), tar (e.g.,Alphosyl, anthralin), topical steroids (e.g., corticosteroids,halobetasol), topical retinoids (e.g., zorac, Tazarotene), systemicantimetabolites (e.g., oral methotrexate), immunosuppressive drugs(e.g., oral cyclosporine, tacrolimus, mycophenolate, and mofetil) andoral retinoids (e.g., acitretin).

Treatment Kits

[0290] In other embodiments, the present invention relates to a kit forconveniently and effectively carrying out the methods in accordance withthe present invention. In general, the pharmaceutical pack or kitcomprises one or more containers filled with one or more of theingredients of the pharmaceutical compositions of the invention. Suchkits are especially suited for the delivery of solid oral forms such astablets or capsules. Such a kit preferably includes a number of unitdosages, and may also include a card having the dosages oriented in theorder of their intended use. If desired, a memory aid can be provided,for example in the form of numbers, letters, or other markings or with acalendar insert, designating the days in the treatment schedule in whichthe dosages can be administered. Alternatively, placebo dosages, orcalcium dietary supplements, either in a form similar to or distinctfrom the dosages of the pharmaceutical compositions, can be included toprovide a kit in which a dosage is taken every day. Optionallyassociated with such container(s) can be a notice in the form prescribedby a governmental agency regulating the manufacture, use or sale ofpharmaceutical products, which notice reflects approval by the agency ofmanufacture, use or sale for human administration.

Equivalents

[0291] The representative examples that follow are intended to helpillustrate the invention, and are not intended to, nor should they beconstrued to, limit the scope of the invention. Indeed, variousmodifications of the invention and many further embodiments thereof, inaddition to those shown and described herein, will become apparent tothose skilled in the art from the full contents of this document,including the examples which follow and the references to the scientificand patent literature cited herein. It should further be appreciatedthat the contents of those cited references are incorporated herein byreference to help illustrate the state of the art.

[0292] The following examples contain important additional information,exemplification and guidance that can be adapted to the practice of thisinvention in its various embodiments and the equivalents thereof.

Exemplification

[0293] The compounds of this invention and their preparation can beunderstood further by the examples that illustrate some of the processesby which these compounds are prepared or used. It will be appreciated,however, that these examples do not limit the invention. Variations ofthe invention, now known or further developed, are considered to fallwithin the scope of the present invention as described herein and ashereinafter claimed.

[0294] 1) General Description of Synthetic Methods:

[0295] The practitioner has a a well-established literature of macrolidechemistry to draw upon, in combination with the information containedherein, for guidance on synthetic strategies, protecting groups, andother materials and methods useful for the synthesis of the compounds ofthis invention.

[0296] The various references cited herein provide helpful backgroundinformation on preparing compounds similar to the inventive compoundsdescribed herein or relevant intermediates, as well as information onformulation, uses, and administration of such compounds which may be ofinterest.

[0297] Moreover, the practitioner is directed to the specific guidanceand examples provided in this document relating to various exemplarycompounds and intermediates thereof.

[0298] The compounds of this invention and their preparation can beunderstood further by the examples that illustrate some of the processesby which these compounds are prepared or used. It will be appreciated,however, that these examples do not limit the invention. Variations ofthe invention, now known or further developed, are considered to fallwithin the scope of the present invention as described herein and ashereinafter claimed.

[0299] According to the present invention, any available techniques canbe used to make or prepare the inventive compounds or compositionsincluding them. For example, a variety of solution phase syntheticmethods such as those discussed in detail below may be used.Alternatively or additionally, the inventive compounds may be preparedusing any of a variety combinatorial techniques, parallel synthesisand/or solid phase synthetic methods known in the art.

[0300] It will be appreciated as described below, that a variety ofinventive compounds can be synthesized according to the methodsdescribed herein. The starting materials and reagents used in preparingthese compounds are either available from commercial suppliers such asAldrich Chemical Company (Milwaukee, Wis.), Bachem (Torrance, Calif.),Sigma (St. Louis, Mo.), or are prepared by methods well known to aperson of ordinary skill in the art following procedures described insuch references as Fieser and Fieser 1991, “Reagents for OrganicSynthesis”, vols 1-17, John Wiley and Sons, New York, N.Y., 1991; Rodd1989 “Chemistry of Carbon Compounds”, vols. 1-5 and supps, ElsevierScience Publishers, 1989; “Organic Reactions”, vols 1-40, John Wiley andSons, New York, N.Y., 1991; March 2001, “Advanced Organic Chemistry”,5th ed. John Wiley and Sons, New York, N.Y.; and Larock 1990,“Comprehensive Organic Transformations: A Guide to Functional GroupPreparations”, 2^(nd) ed. VCH Publishers. These schemes are merelyillustrative of some methods by which the compounds of this inventioncan be synthesized, and various modifications to these schemes can bemade and will be suggested to a person of ordinary skill in the arthaving regard to this disclosure.

[0301] The starting materials, intermediates, and compounds of thisinvention may be isolated and purified using conventional techniques,including filtration, distillation, crystallization, chromatography, andthe like. They may be characterized using conventional methods,including physical constants and spectral data.

[0302] Certain exemplary compounds of the invention are listed below andare referred to by compound number as indicated. B2193

B2194

B2215

B2292

B2293

B2297

B2329

B2331

B2337

B2338

B2356

B2357

B2358

B2359

B2366 & B2365

B2395

B2396

B2397

B2500

B2501

single enantimer B2522

B2526

B2538

B2543

racemic diastereomer 1 B2544

racemic diastereomer 2 B2545

Racemic isomer 1 ER803026

ER803029

ER803030

ER803064

ER803591

diastereomer of ER803593 ER803593

diastereomer of ER803591 ER803604

ER803734

ER803758

ER803829

ER803882

Stereochemistry @ C4 needs confirmation ER803916

ER803918

mixture of products ER803924

ER804003

ER804018

ER804019

ER804022

ER804035

ER804060

ER804103

ER804104

ER804131

ER804142

ER804143

ER804168

ER804189

ER804387

ER804401

ER804428

ER804446

ER804504

ER804505

ER804555

ER804556

ER804567

ER804584

ER804595

ER804606

ER804622

ER804630

ER804710

ER804730

ER804731

ER804734

ER804744

ER804745

ER804746

ER804747

ER804755 & ER804756

ER804758

ER804759

ER804778

ER804779

ER804784

ER804793

ER804863

ER804986

ER805023

ER805053

ER805125

ER805135

ER805146

ER805149

ER805189

ER805190

ER805192

ER805215

ER805216

ER805217

ER805218

ER805221

ER805223

ER805227

ER805228

ER805229

ER805232

ER805233

ER805709

ER805855

ER805882

ER805911

ER805940

ER805977

ER805998

ER806201

ER806203

ER806204

ER806328

ER806563

ER806621

ER806624

ER806752

ER806776

ER806795

ER806821

ER806907

ER807209

ER807551

ER807563

ER808064

ER808129

ER890003

ER890004

ER890005

ER890006

ER890007

ER890008

ER890009

F152acetonite

NF0530

NF0531

NF0552

NF0675

NF0761

C10 stereochemistry undefined NF0879

NF0880

NF0887

NF0905

NF0934

BF1226 and NF1227

NF1418

NF1419

NF1535

NF1537

cis/trans, 1:10 NF1774

NF1872

NF2306

NF2432

NF2433

NF2435

NF2436

NF2544

NF2545

NF2546

NF2547

NF2548

NF2550

NF2551

NF2552

NF2553

NF2554

NF2555

NF2556

NF2557

NF2558

NF2559

NF2560

NF2561

[0303] General Reaction Procedures:

[0304] Unless mentioned specifically, reaction mixtures were stirredusing a magnetically driven stirrer bar. An inert atmosphere refers toeither dry argon or dry nitrogen. Reactions were monitored either bythin layer chromatography, by proton nuclear magnetic resonance (NMR) orby high-pressure liquid chromatography (HPLC), of a suitably worked upsample of the reaction mixture.

[0305] Listed below are abbreviations used for some common organicreagents referred to herein: m-CPBA: meta-chloroperbenzoic acid DDQ:2,3-Dichloro-5,6-dicyano-1,4-benzoquinone DEAD Diethyl azodicarboxylateDIBAL-H: Diisobutyl aluminum hydride DMAP: N,N-DimethylaminopyridineDMF: N,N-Dimethylformamide HMPA: Hexamethylphosphoramide LDA: Lithiumdiisopropyl amide LiHMDS: Lithium bis (trimethylsilyl) amide PCC:Pyridinium chlorochromate TBAF: Tetrabutylammonium fluoride THF:Tetrahydrofuran

[0306] General Work Up Procedures:

[0307] Unless mentioned specifically, reaction mixtures were cooled toroom temperature or below then quenched, when necessary, with eitherwater or a saturated aqueous solution of ammonium chloride. Desiredproducts were extracted by partitioning between water and a suitablewater-immiscible solvent (e.g. ethyl acetate, dichloromethane, diethylether). The desired product containing extracts were washedappropriately with water followed by a saturated solution of brine. Onoccasions where the product containing extract was deemed to containresidual oxidants, the extract was washed with a 10% solution of sodiumsulphite in saturated aqueous sodium bicarbonate solution, prior to theaforementioned washing procedure. On occasions where the productcontaining extract was deemed to contain residual acids, the extract waswashed with saturated aqueous sodium bicarbonate solution, prior to theaforementioned washing procedure (except in those cases where thedesired product itself had acidic character). On occasions where theproduct containing extract was deemed to contain residual bases, theextract was washed with 10% aqueous citric acid solution, prior to theaforementioned washing procedure (except in those cases where thedesired product itself had basic character). Post washing, the desiredproduct containing extracts were dried over anhydrous magnesiumsulphate, and then filtered. The crude products were then isolated byremoval of solvent(s) by rotary evaporation under reduced pressure, atan appropriate temperature (generally less than 45° C.).

[0308] On occasions where triphenylphosphine oxide was a major byproductof the reaction, the reaction mixture was added directly to a largevolume of well-stirred hexane. The resultant precipitate oftriphenylphosphine oxide was removed by filtration and the filtrateprocessed in the usual manner.

[0309] General Purification Procedures:

[0310] Unless mentioned specifically, chromatographic purificationrefers to flash column chromatography on silica, using a single solventor mixed solvent as eluent. Suitably purified desired product containingelutes were combined and concentrated under reduced pressure at anappropriate temperature (generally less than 45° C.) to constant mass.Final compounds were dissolved in 50% aqueous acetonitrile, filtered andtransferred to vials, then freeze-dried under high vacuum beforesubmission for biological testing.

[0311] Synthesis for commonly Used Intermediates:

[0312] Starting material (50.0 g, 0.27 mol) was dissolved in 650 mL ofTHF at 0° C. Triphenylphosphine (93.6 g, 0.35 mol) was added, followedby methanol (12.2 mL, 0.30 mol) and diethyl azodicarboxylate (56.2 ml,0.35 mol). After stirring at 0° C. for 1.5 h, the reaction mixture wasconcentrated, redissolved in diethyl ether, washed with 1N sodiumhydroxide solution. The aqueous layer was acidified with concentratedhydrochloric acid and extracted with diethyl ether. After purificationon silica gel column, 42.0 g of 509-HD-207 was obtained as a pale yellowsolid in 78% yield.

[0313] To the reaction flask containing NaH (95%, 14.5 g, 0.57 mol) in 1L of THF at 0° C. was added 509-HD-207 (75.0 g, 0.38 mol) in 0.5 L ofTHF. After stirred for 0.5 h, chloromethyl methyl ether (43.6 mL, 0.57mol) was added. After stirred at 0° C. for 1 h, it was warmed up to roomtemperature. The reaction was quenched with water and extracted withpentane. After purification on silica gel column, 83 g of 509-HD-209 wasobtained as colorless oil in 92% yield.

[0314] Diisopropyl amine (68.1 mL, 486 mol) was dissolved in 1 L of THFat 0° C. n-BuLi (2.5 M, 207 mL) was added. The solution was stirred for20 min, and then cooled down to −78° C. The solution of 509-HD-209 (77.8g, 324 mol) in 250 mL of THF was added slowly. 1 h later the solution ofdiphenyl diselenide (85.9 g, 275 mol) in 250 ml of THF was added. Afterstirring at −78° C. for 1 h, the reaction was quenched with saturatedammonium chloride solution, and extracted with diethyl ether. Afterpurification on silica gel column, 90.2 g of 509-HD-211 was obtained aspale yellow oil in 68% yield.

[0315] 509-HD-211 (90.2 g, 228 mmol) was dissolved in 500 mL of ethanol.Sodium hydroxide solution (1N, 456 mL) was added. The resulting solutionwas heated under reflux for 12 h. The reaction mixture was acidifiedwith 1N hydrochloric acid, extracted with diethyl ether andconcentrated, giving 84.6 g of 509-HD-212 as a pale yellow solid in 97%yield.

[0316] 509-HD-212 (84.6 g, 222 mmol) and triphenylphosphine (75.7 g, 289mmol) was dissolved in a mixture of 500 mL of diethyl ether and 125 mLof toluene at 0° C. 2-(trimethylsilyl) ethanol (38.2 mL, 266 mmol) anddiethyl azodicarboxylate (45.4 mL, 289 mmol) were added respectively.After stirred for 10 min, it was warmed up to room temperature. Largeamount of pentane was added to precipitate the solid. After filtration,the crude product was purified on silica gel column and 80.0 g of509-HD-213 was obtained as pale yellow oil in 75% yield.

[0317] To a solution of starting material (157 g, 0.86 mol) in 1.6 L oftoluene (slightly cloudy), TMS-ethanol (150 g, 1.27 mol, 1.48 eq.), andPPh₃ (440 g, 1.68 mol, 1.95 eq.) were added. After cooled to 0° C., DEAD(725 mL of 40% solution, 1.29 mol, 1.5 eq.) was slowly added by droppingfunnel while maintaining internal temp below 10° C. in three hours.After stirred at rt for 48 h, it was poured into a rapid stirredsolution of hexanes (12 L). The solid was filtered through a celite padand the pad was washed with 1 L of hexanes. The filtrates were combinedand concentrated to give the crude product as brown oil. The oil waspurified on silica gel with hexanes/EtOAc (15:1, 10:1, 6:1) to give 140g of product as an off white solid.

[0318] The phenol (140 g, 0.49 mol) was dissolved in 400 mL of CH₂Cl₂,DBU (135.5 mL, 0.91 mol, 1.85 eq.) was added. After cooled to 0° C.,MOMCl (66 mL, 0.87 mol, 1.78 eq.) was added. After stirred at rt for 24h, it was quenched with Sat. NH₄Cl, extracted with EtOAc. The organiclayer was washed with brine, dried and concentrated to dryness. Thecrude product was purified on column with hexanes/EtOAc, 10:1, 6:1 togive 132 g of desired product (82% in yield).

[0319] To a solution of diisopropylamine (143.6 mL, 1.02 mol, 2.3 eq.)in 320 mL of THF, n-BuLi (422.6 mL, 2.5 M) was added at 0° C. byaddition funnel, while controlling the internal temperature around 5° C.After 5 min at that temperature, the reaction was cooled to −78° C. Asolution of starting material (145 g, 0.44 mol) in 475 mL of THF wasadded by addition funnel while internal temperature controlled at orbelow −70° C. After addition, it was stirred at −70° C. for 30 min. Thenat that temperature, a solution of PhSe₂ (140 g, 0.45 mol, 1 eq.) in 400mL of THF was added by addition funnel. After addition, it was stirredat −78° C. for 45 min. The reaction was quenched with sat. NH₄Cl,diluted with EtOAc. It was warmed to rt. The organic layer was washedwith brine, dried and concentrated to dryness. The crude product wasused without purification for next step.

[0320] The crude product from last step was dissolved in 300 mL of EtOH.Then 300 mL of 1N NaOH was added. The reaction was heated at 80° C.overnight. After cooled, it was transferred to a separatory funnel andwashed with hexanes. The aq. Layer was acidified at 0° C. with 1N HCl topH=3. Then it was extracted with EtOAc (3×). The combined organic layerswere washed with brine, dried and concentrated to dryness. The crudeacid was used for next step without purification.

[0321] The esterification was run according to the first step using DEAD(200 g), PPh₃ (330 g) and TMS-ethanol (150 g) in 1.4 L of toluene togive 145 g of product after column chromatography.

[0322] Other C14-substitutions aromatic pieces such as methyl, ethyl,Benzyl, PMB (MPM) etc. were made in analogues manner by substituting thefirst step with corresponding alcohols such as methanol, ethanol, benzylalcohol, or PMB alcohol etc.

[0323] To a stirring solution of diisopropylamine (2 eq., 366 mmol, 51.3mL) in dry THF (200 mL) at −78° C. was added slowly a 1.6 M solution ofn-BuLi (2 eq., 366 mmol, 230 mL) over a period of 20 min. The reactionmixture was warmed to 0° C. and allowed to stir for 45 min after whichthe solution was cooled back to −78° C. Then, a solution ofmethyl-3-hydroxybutyrate (21.6 g, 183 mmol) in dry THF (100 mL) wasadded slowly over a period of 20 min after which neat Mel (Seq, 915mmol, 57 mL) was added over a period of 5 min. The reaction mixture wasallowed to stir for 10 min at −78° C. then warmed to rt, and stirred for2 h. The reaction was quenched with a saturated solution of NH₄Cl (350mL), extracted with Et₂O (3×400 mL), the combined organic extracts werewashed with a saturated solution of NH₄Cl (350 mL), water (2×450 mL),brine (450 mL), dried with K₂CO₃, filtered and concentrated. The crudealcohol 555-RB-224 was dissolved in dry DMF (100 mL), imidazole (2.5 eq,363 mmol, 24.7 g) was added and the mixture was cooled to 0° C. inice/water bath. Then TBSCI (1.2 eq, 33.0 mmol, 5 g) was added, themixture was allowed to warm slowly to rt and stirred for 16 h afterwhich a saturated solution of NaHCO₃ (250 mL) was added. The mixture wasextracted with Et₂O (3×250 mL) and the combined organic extracts werewashed with a saturated solution of NaHCO₃ (350 mL), water (3×350 mL),brine (350 mL), dried with Na₂SO₄, filtered and concentrated. The crudeproduct was purified by chromatography on silica gel using 5%EtOAc/hexane to give 31.7 g (129 mmol, 70% 2 steps) of the protectedcompound 554-RB-225.

[0324] In a 5 L three neck flask equipped with mechanical stirring wasplaced 554-RB-225 (221 mmol, 54.5 g) dissolved in toluene (750 mL). Themixture was cooled to −78° C. and DIBAL-H (1M in toluene, 2.5 eq., 553mmol, 553 mL) was added slowly. The mixture was allowed to stir at −78°C. for 15 min after which it was warmed to 0° C. and stirred for 2 h.Reaction was quenched with MeOH (10 eq., 2.2 mol, 89 mL) at −78° C. andallowed to warm to rt, Et₂O (2.5 L) and a saturated solution of Na₂SO₄(1 L) were added and the solution was stirred overnight. Mixture wasfiltered through celite and the solid was washed with Et₂O (2×1 L). Thefiltrate was concentrated under reduced pressure and the resultingresidue was purified by chromatography on silica gel using 10-15%EtOAc/hexane to give 40.6 g (186 mmol, 76%) of alcohol 554-RB-227.

[0325] To a solution of oxalyl chloride (2 eq., 372 mmol, 33.0 mL) inCH₂Cl₂ (800 mL), DMSO (4 eq., 744 mmol, 53.0 mL) was added at −78° C.After 30 min at −78° C., a solution of alcohol 554-RB-227 (186 mmol,40.6 g) in CH₂Cl₂ (200 mL) was added over a period of 15 min. After 50min at −78° C., Et₃N (4 eq., 744 mmol, 104 mL) was added and thereaction was warmed to 0° C. and stirred for 45 min. It was quenchedwith a saturated solution of NH₄Cl (500 mL), extracted with EtOAc (1×2L, 2×400 mL). The combined organic layers were dried with Na₂SO₄,filtered and concentrated. The crude aldehyde was filtered through ashort silica gel column with 10% EtOAc/hexane. To a solution of PPh₃ (3eq., 558 mmol, 146 g) in CH₂Cl₂ (2 L), CBr₄ (1.5 eq., 279 mmol, 92.5 g)was added at 0° C. Then, a solution of aldehyde and Et₃N (1 eq., 186mmol, 26 mL) in CH₂Cl₂ (200 mL) was added. The solution was stirredunder nitrogen at rt for one hour after which the mixture wasconcentrated under reduced pressure. The residue was dissolved inCH₂Cl₂, poured into hexane (2.5 L) and stirred. The precipitate wasfiltered through celite and the filtrate was concentrated. Purificationby chromatography on silica gel using CH₂Cl₂ as eluent gave 62.9 g (169mmol, 91% 2 steps) of 554-RB-228.

[0326] A 5 L, 3-neck flask was equipped with mechanical stirring,cooling bath and flushed with nitrogen. Then, a solution of alcohol491-HAD-46 (202 mmol, 52.5 g) and MPMOTCI (2 eq., 404 mmol, 115.5 g) inEt₂O (1 L) was added to the flask and cooled to 0° C. A solution of TfOH(1.5 mL) in Et₂O (120 mL) was added slowly with a syringe pump over aperiod of 50 min. Then, a saturated solution of NaHCO₃ (500 mL) wasadded, the mixture was extracted with Et₂O (2×700 mL); the combinedorganic extracts were washed with brine (2×1 L), dried over Na₂SO₄ andconcentrated. The residue was dissolved in CH₂Cl₂, poured in hexane (2.5L), the precipitated was filtered through celite and the filtrate wasconcentrated under reduced pressure. The crude material was purified bychromatography on silica gel using 5-10% EtOAc/hexane as eluent to give57.3 g (151 mmol, 75%) of the protected alcohol 554-RB-235.

[0327] In a 3 L, three neck flask equipped with mechanical stirring wasplaced 554-RB-235 (151 mmol, 57.3 g) dissolved in toluene (750 mL). Themixture was cooled to −78° C. and DIBAL-H (1M in toluene, 2.65 eq., 400mmol, 400 mL) was added slowly. The mixture was allowed to stir at −78°C. for 10 min after which it was warmed to 0° C. and stirred for 20 min.Reaction was quenched with MeOH (10 eq., 1.5 mol, 61 mL) at −78° C. andallowed to warm to rt. Et₂O (2.5 L) and a saturated solution of Na₂SO₄(1 L) were added and the solution was stirred overnight. Mixture wasfiltered through celite and the solid was washed with Et₂O (2×1 L). Thefiltrate was concentrated under reduced pressure and the resultingresidue was purified by chromatography on silica gel using 20-40%EtOAc/hexane to give 27 g (91 mmol, 60%) of alcohol 554-RB-237.

[0328] To a solution of oxalyl chloride (2 eq., 202 mmol, 18 mL) inCH₂Cl₂ (650 mL), DMSO (4 eq., 404 mmol, 29 mL) was added at −78° C.After 30 min at −78° C., a solution of alcohol 554-RB-237 (101 mmol, 30g) in CH₂Cl₂ (100 mL) was added over a period of 30 min. After 45 min at−78° C., Et₃N (4 eq., 404 mmol, 56 mL) was added and the reaction waswarmed to 0° C. and stirred for 45 min. It was quenched with a saturatedsolution of NH₄Cl (250 mL), extracted with EtOAc (1×2 L, 2×250 mL). Thecombined organic layers were dried with Na₂SO₄, filtered andconcentrated. The crude aldehyde was purified by chromatography onsilica gel with 10% EtOAc/hexane to give 27 g (91.7 mmol, 91%) ofaldehyde 554-RB-238.

[0329] Dibromoolefin 554-RB-228 (1.5 eq., 138 mmol, 51.2 g) wasdissolved in THF (1 L) and cooled to −78° C., under nitrogen. Then,n-BuLi (1.6M/hexane, 3.3 eq., 302 mmol, 189 mL) was added and thereaction was stirred at −78° C. for 40 min, at 0° C. for 30 min, thencooled back to −78° C. Aldehyde 554-RB-238 (91.7 mmol, 27.0 g) dissolvedin THF (200 mL) was added to the solution and stirred for 30 min at −78°C. The solution was allowed to warm to rt and was stirred for 1.5 hrs.The mixture was quenched with water (700 mL), extracted with EtOAc(3×750 mL) and the combined organic extracts were washed with brine (1L), dried with Na₂SO₄, filtered and concentrated. The residue waspurified by chromatography on silica gel using 10-30% EtOAc/hexane togive 43.7 g (86 mmol, 94%) of 554-RB-240.

[0330] 554-RB-240 (86 mmol, 43.7 g) was dissolved in hexane (1 L). Then,quinoline (1 mL) and Lindlar catalyst (10 g) were added. H₂ balloon wasmounted and the mixture was purged 5× with H₂. Reaction was stirredunder hydrogen. After 13 hrs, 17 hrs, 22 hrs, and 38 hrs, catalyst wasfiltered and new catalyst (10 g) and quinoline (1 mL) were added eachtime. After 42 hrs, reaction was stopped, catalyst was filtered throughcelite and mixture was concentrated under reduced pressure. Then, crude554-RB-241 was dissolved in CH₂Cl₂ (700 mL), Et₃N (3.75 eq., 323 mmol,45 mL), BzCl (3 eq., 258 mmol, 30 mL) and DMAP (0.075 eq., 6.45 mmol,788 mg) were added and the mixture was stirred for 96 hrs at rt undernitrogen. The mixture was diluted with EtOAc (2 L) and a 0.1N solutionof NaOH (800 mL). Organic layer was separated and the aqueous phase wasextracted with EtOAc (2×500 mL). The organic combined extracts werewashed with a 0.2N solution of NaOH (5×500 mL), brine, dried withNa₂SO₄, filtered and concentrated. The crude compound was filteredthrough a silica gel column with 5% EtOAc/hexane to give a quantitativeyield of protected compound 554-RB-242.

[0331] 554-RB-242 was dissolved in CH₂Cl₂ (500 mL), H₂O (250 mL) and DDQ(1.1 eq., 94.6 mmol, 21.3 g) were added and the mixture was stirredvigorously at rt for 4 hrs. The mixture was quenched with a 0.2Nsolution of NaOH (500 mL) and diluted with EtOAc (2 L). The organiclayer was separated and the aqueous phase was back extracted with EtOAc(2×500 mL). The combined organic layers were washed with a 0.2N solutionof NaOH (3×700 mL), brine (700 mL), dried with Na₂SO₄, filtered andconcentrated. The crude residue was purified by chromatography on silicagel using 10% EtOAc/hexane to give 39.9 g (81 mmol, 94% 3 steps fromacetylene) of free alcohol 554-RB-244.

[0332] To a solution of 554-RB-244 (6.09 mmol, 3.0 g) in toluene (100mL), Ph₃P (1.7 eq., 10.4 mmol, 2.71 g) was added at rt. Then, CH₃I (1.3eq., 7.92 mmol, 0.49 mL) and DEAD (1.1 eq., 6.70 mmol, 1.45 mL) wereadded at the same time. The mixture was stirred for 1.5 hrs at rt afterwhich it was poured in hexane and stirred for 10 min. The precipitatewas filtered and the filtrate was concentrated. The residue was purifiedby chromatography on silica gel using 5% EtOAc/hexane to give 3.40 g(5.64 mmol, 93%) of iodide 554-RB-260.

[0333] Preparation of C4-H Series Acyclic Segment:

[0334] To a solution of 531-YW-2-3 (7.5 g) in 20 mL of DMF, imidazole (5g) and TBDPSCl (8.4 g) were added. During the addition exotherm wasobserved. After 3 h, it was diluted with EtOAc, washed with aq. Sat.NH₄Cl and brine. After drying and filtration, it was concentrated. Thecrude product was purified on silica gel with Hexanes/EtOAc, 20:1, and10:1 to give 11.0 g of desired product.

[0335] The product was dissolved in 200 mL of THF. LAH (1.4 g) was addedat 0° C. After 10 min at 0° C., it was quenched with water, 1N NaOH. Themixture was stirred at rt for 1 h. Then it was filtered. The combinedfiltrates were concentrated to dryness. The crude product was purifiedon silica gel with Hexanes/EtOAc, 10:1, 4:1, and 2:1 to give 9.0 g ofdesired product with satisfactory 1H NMR spectra, 343-YW-275.

[0336] To a solution of commercial available (s)-3-hydroxy butanol (10g, Aldrich) in 50 mL of DMF, TsOH (20 mg, catalytic) and MeOPhCH(OMe)₂(24 g) were added. After 3 h at 35° C. on a rotovap with slight vacuum,it was cooled and quenched with aq. Sat. NaHCO₃. The mixture wasextracted with EtOAc (3×). The organic layers were washed with brine(2×), dried and concentrated. The crude product was evaporated withtoluene (3×).

[0337] The crude product was dissolved in 700 mL of CH₂Cl₂. At 0° C.,DIBAL-H solution (200 mL, 1.0 M, excess) was added. The reaction waswarmed to room temperature overnight. Then it was quenched with methanol(50 mL), sat. Na₂SO₄ at 0° C. The mixture was diluted with Et₂O (1.5 L).After stirred for 5 h, it was filtered through a pad of celite. Thefiltrate was concentrated to give an oil. The oil was purified on silicagel with Hexanes/EtOAc, 10:1, 6:1, 3:1, and 1:1 to give 24 g of desiredproduct, 343-YW-203.

[0338] To a solution of DMSO (3.7 mL) in 150 mL of CH₂Cl₂, a solution of343-YW-203 (3.6 g) in 50 mL of CH₂Cl₂ was added. At 0° C., solid P₂O₅(6.06 g) was added. was warmed to room temperature. After stirred atroom temperature for 1 h, the reaction turned light pink and was cooledto 0° C. Et₃N (12 mL) was added. After 15 min at 0° C., it was warmed toroom temperature. After 10 min, it was quenched with sat. NH₄Cl, andextracted with CH₂Cl₂ (2×). The organic layers were dried andconcentrated. The crude product was suspended in Et₂O and filtered. Thefiltrates were concentrated to dryness.

[0339] To a solution of PPh₃ (11.6 g) in 30 mL of CH₂Cl₂, CBr₄ (7.4 g)was added at 0° C. The internal temperature was controlled below 10° C.After 10 min, a solution of aldehyde in 20 mL of CH₂Cl₂ was added. Theinternal temperature went up to 20° C. It was warmed to room temperatureand stirred for 1h. Then it was poured into a rapid stirring pentanesolution. The precipitation was filtered. The filtrates wereconcentrated. The crude product was purified on silica gel column withhexanes/EtOAc, 20:1, 15:1, 10:1 to give 4.4 g of the desired product,343-YW-276.

[0340] Oxidation of alcohol: To a solution of 343-YW-275 (3.6 g) in 25mL of CH₂Cl₂, DMSO (1.84 mL) was added followed by P₂O₅ (3.65 g) solidat 0° C. Then it was warmed to room temperature for 1 h. After cooled to0° C., Et₃N (5.94 mL) was added. After stirred at 0° C. for 30 min, itwas warmed to room temperature. After stirred at room temperature for 3h, it was quenched with sat. NH₄Cl, and extracted with CH₂Cl₂ (2×). Theorganic layers were washed with brine, dried and concentrated. The crudeproduct was purified on silica gel with Hexanes/EtOAc, 10:1, 6:1, and4:1 to give 2.5 g of aldehyde, 343-YW-277.

[0341] Coupling: To a solution of 343-YW-276 (4.5 g, 12.36 mmol, 2 eq.)in 35 mL of THF, n-BuLi (5.4 mL, 2.5M, 2.2 eq.) was added at −78° C.After 10 min at −78° C., it was warmed to room temperature for 30 min.After cooled back to −78° C., a solution of 343-YW-277 (2.5 g, 6.06mmol, 1 eq.) in 10 mL of THF was added. It was then warmed to 0° C.,after 30 min. After 4 h at 0° C., it was quenched with aq. Sat. NH₄Cl,extracted with EtOAc (2×). The organic layers were washed with brine,dried and concentrated. The crude product was purified on silica gelwith Hexanes/EtOAc, 20:1, 15:1, 10:1, 6:1 to give 2.5 g of desiredproduct, 343-YW-278 along with recovered 343-YW-276.

[0342] To a solution of 343-YW-278 (2.50 g) in 100 mL of hexanes,Lindlar catalyst (330 mg, catalytic) and quinoline (50 μL, catalytic)were added. After degassed under vacuum and refilled with H₂ for severaltimes, it was stirred under hydrogen balloon for 3 h. Then the catalystwas filtered and fresh catalyst was added. After degassing, it wasstirred under hydrogen overnight. The reaction was filtered throughcelite. The filtrates were combined and concentrated to dryness to give2.4 g of desired product, 343-YW-279.

[0343] To a solution of 343-YW-279 (2.4 g) in 15 mL of CH₂Cl₂, BzCl (0.9mL), Et₃N (2 mL) and DMAP (50 mg) were added. After 18 h, another 200 uLof BzCl was added. After total 24 h, it was quenched with aq. Sat. NH₄Cland extracted with EtOAc (2×). The organic layers were washed withbrine, dried and concentrated. The crude product was purified on silicagel with Hexanes/EtOAc, 20:1, 10:1, and 6:1 to give 2.2 g of desiredester.

[0344] To a solution of the ester from last step in 10 mL of THF, solidTBAF was added. After 18 h, it was quenched with sat. NH₄Cl andextracted with EtOAc. The organic layer was washed with brine, dried andconcentrated. The crude product was purified on silica gel withHexanes/EtOAc, 10:1, 6:1, 4:1, and 2:1 to give 1.45 g of alcohol.

[0345] To a solution of the alcohol from last step and PPh₃ (1.3 g) in20 mL of toluene, DEAD (750 μL) and Mel (250 μL) were addedsimultaneously at room temperature. After 30 min, it was diluted withCH₂Cl₂ to a clear solution. Then it was poured into pentanes with rapidstirring. The precipitation was filtered through a celite pad. Thefiltrate was concentrated. The crude product was purified on silica gelwith Hexanes/EtOAc, 20:1, 10:1, 8:1 to give 1.5 g of desired product,343-YW-281. A satisfactory ¹H NMR was obtained.

[0346] Preparation of Advance Phenol Intermediate for C14-AnalogSynthesis:

[0347] Iodide 2 (4.94 g, 8.2 mmol, 1.0 eq.) and selenide 3 (7.00 g, 12.3mmol, 1.5 eq.) were dissolved in HMPA (10.0 mL) and THF (90 mL). Theresulting mixture was magnetically stirred at −78° C. and slowly addedover 50 min LiHMDS (18.0 mL, 9.0 mmol, 1.1 eq., LiHMDS 0.5M in THF,addition rate of 0.32 mL/min). The reaction mixture was stirred 1 h and45 min at −78° C., quenched with NH₄Cl sat. (200 mL), diluted with H₂O,added ethyl acetate (500 mL). The layers were separated and the aqueousone was extracted with ethyl acetate (2×500 mL). The combined organiclayers were washed with H₂O (2×300 mL), dried with Na₂SO₄, filtered, andconcentrated under reduced pressure. The crude oil was purified on aSiO₂ column (230-400 Mesh silica). The products were dissolved in hexaneprior to be loaded on the column. Elution: 3%, then 5% ethylacetate/hexane. The desired product 4 (5.43 g, 64% yield) was isolatedas viscous brownish oil.

[0348] The crude substrate 4 (mixture of selenide 3 and coupled material4, <58.9 mmol) was dissolved in CH₂Cl₂ (750 mL), cooled down to 0° C.and added in small portions MCPBA (Aldrich 57-86%, 43.5 g, >144 mmol,2.4 eq.).

[0349] The first portions are exothermic and towards the end of theaddition, no exotherm was noticed, Tmax was 4° C. Stirred for 45 min at0° C., triethylamine (50 mL, 357 mmol, 6 eq.) was added SLOWLY!EXOTHERMIC! Tmax=10° C. Once the exotherm had ceased, the reactionmixture was warmed up to rt. Stirred for 60 min at that temperature, asolution of NaS₂O₃ (51.5 g) was prepared using NaHCO₃ saturated anddistilled water. Layers were separated; the aqueous one was extractedtwice with CH₂Cl₂. The combined organic layers were dried with Na₂SO₄,filtered, concentrated down, added crude material from a previoussmall-scale run, purified on a SiO₂ column (1.25 kg of 230-400 Meshsilica from Silicycle). The crude material was loaded on the column as aslurry prepared with 3% ethyl acetate/hexane and 230-400 Mesh silica.Elution: 3% (8 L), 7.5% (8 L) and 10% (6 L) ethyl acetate/hexane. Thedesired material 5 (16.3 g, 30% combined yield since the selenidecoupling) is viscous oil.

[0350] Substrate 5 was dissolved in THF (43 mL), added an imidazole.HClbuffered TBAF solution (60.5 mL, 60.5 mmol of TBAF, 3.4 eq. of TBAF 1 Min THF and 45 mmol of imidazole.HCl, 2.5 eq. of imidazole.HCl). Thatbuffered solution was prepared as follows: imidazole.HCl was dissolvedin a commercial 1 M TBAF/THF solution to give a resulting imidazole.HClmolarity of 0.75 M. The resulting reaction mixture was stirred 2 min atrt then it was added drop-wisely a regular TBAF solution (76 mL, 76mmol, TBAF 1.0 M in THF). The reaction mixture was stirred in an oilbath at 50° C. during a total of 88 h, cooled down to rt, added NH₄Clsat. (300 mL) and Et₂O (300 mL). The layers were separated and theaqueous was extracted with Et₂O (3×150 mL). The organic layers werecombined, washed with brine (3×100 mL), dried with Na₂SO₄, filtered,concentrated to dryness, azeotroped twice with Et₂O (2×100 mL) givingdesired 6 that was used crude for the next step.

[0351] In a three neck 5 L flask equipped with a condenser and anaddition funnel, was added CH₂Cl₂ (2 L) followed by triethylamine (7.6mL, 54.0 mmol, 3.0 eq.) and 2-chloro-1-methyl pyridinium iodide (14.2 g,54.0 mmol, 3.0 eq.). The resulting mixture was warmed up to reflux andadded drop-wisely a CH₂Cl₂ solution of hydroxy-acid 6 (14.4 g of crudematerial in 230 mL of CH₂Cl₂). The addition took 3 h and the resultingreaction mixture was stirred at reflux for 12 h, then cooled down to rt,salts were filtered and CH₂Cl₂ was removed under reduced pressure. Theresidue was dissolved in Et₂O, washed with a 1:1 mixture of saturatedbrine and saturated NaHCO₃. The aqueous layer was extracted twice withEt₂O. The combined organic layers were washed with brine, dried withNa₂SO₄ and concentrated under reduced pressure, purified on a SiO₂column (250 g of 230-400 Mesh silica from Silicycle). The crude materialwas dissolved in CH₂Cl₂ prior to be loaded on the column. Elution: 15%,25%, 35% ethyl acetate/hexane. The desired macrocycle 7 (7 g) was stillslightly contaminated was used directly for the next step.

[0352] To a solution of 7 (7 g, <18 mmol, 1.0 eq.) in THF (40 mL) at rt,was added drop-wisely TBAF (85 mL, 85 mmol, 4.7 eq., TBAF 1 M in THF).The reaction mixture was stirred at rt for 3 h, and then quenched withNH₄Cl sat. (250 mL) and Et₂O (250 mL). The layers were separated and theaqueous was extracted with Et₂O (2×150 mL). The combined organic layerswere washed with brine (2×100 mL), dried with Na₂SO₄ and concentratedunder reduced pressure, purified on a SiO₂ column (75 g of 230-400 Meshsilica from Silicycle). The crude material was dissolved in CH₂Cl₂(15-20 mL) prior to be loaded on the column. The column was preparedusing 25% ethyl acetate/hexane. Elution: 25%, 35%, 50% ethylacetate/hexane. The desired material 8 (5.10 g, 50% combined yield from5) is a white foam.

[0353] Preparation of Intermediate for C3-C4 Modification Series:

[0354] To the stirred suspension of 2-deoxy-D-ribose (100.8 g, 0.75 mol,commercially available from TCI) in EtOAc (800 ml), were added2-methoxypropene (94 ml, 0.98 mol, 1.3 eq., Aldrich) and PPTS (4.16 g,17 mmol, 2 mol%, Aldrich) at room temperature under N₂. The mixture wasstirred vigorously for 3 hr.

[0355] Then the insoluble residue (remained SM) was filtrated out andTEA (4.6 ml, 2 eq. to PPTS) was added to the filtrate. The resultantfiltrate was concentrated under reduced pressure and the residue waspurified by silica gel flash column chromatography (silica gel 4 kg,hexane-EtOAc 9:1 to 1:1 as eluent) to give 68 g of desired compound ascolorless oil (52%).

[0356] To the stirred suspension of LiAlH₄ (9.26 g, 0.244 mol, 1.25 eq., Wako) in THF (200 mL) cooled in ice/brine bath, was added drop wiseSM (68 g, 0.39 mol) in THF (600 mL+100 mL rinse) under N₂ (in ca. 1.5hr). Then the mixture was stirred for additional 15 min. After quenchingby careful addition of MeOH, 9.26 ml of water, 9.26 ml of 10% NaOH aq.,27.78 mL of water were added successively and the mixture was stirredvigorously for 1 hr. Then, the insoluble material was filtered out usingCelite and washed with EtOAc (500 mL×4), and the resultant filtrate wasconcentrated under reduced pressure, dried in vacuo to give 62.23 g ofthe crude product as light yellow oil (90.5%).

[0357] To the stirred suspension of NaH (8.09 g, 60% oil dispersion, 202mmol, 2.2 eq., Wako) in DMF (200 mL) cooled in ice/brine bath, was addeddrop wise (16.2 g, 91.9 mmol) in DMF (500 mL+100 mL rinse) under N₂. Theresultant mixture was stirred for 75 min. at room temperature. Then themixture was cooled to −55° C. (inner temp.)**, PivCl (12.5 mL, 102 mmol,1.1 eq., TCI) was added drop wise (in ca 10 min). After addition, themixture was allowed to warm to −30° C.

[0358] Quenching was performed by careful addition of sat. NH₄Cl aq.,then the mixture was extracted with EtOAc (1 L). After re-extraction ofthe aqueous layer with EtOAc (500 mL), the combined organic phase waswashed with water (0.6 L×3), brine (0.3 L) and dried over anhydrousNa₂SO₄. After filtration of drying agent, the filtrate was concentratedand the residual brown oil (25.43 g) was purified by silica gel flashcolumn chromatography (silica gel 2.8 kg, hexane-EtOAc 2:1 to 1:1 aseluent) to give 3.73 g of less polar undesired protected mono-ol,531-YW-2-2 (16%) and 13.14 g of polar desired product, 531-YW-2-3 (55%),respectively as colorless oil.

[0359] Iodide Formation:

[0360] To a solution of 531-YW-2-3 (9.5 g, 36.5 mmol) in 400 mL oftoluene, PPh₃ (18 g, 62.1 mmol, 1.9 eq.) was added. Then Mel (2.94 mL,47 mmol, 1.3 eq.) and DEAD (6.29 mL) were added simultaneously by twosyringe-pumps in 20 min. After stirred at room temp for 20 min, it waspoured into a rapid stirred pentane solution. The solid was dissolved bysmall amount of CH₂Cl₂ and added into the pentane. The precipitation wasfiltered through celite, the pad was washed with pentane. The combinedfiltrates were concentrated. The crude oil was purified quickly on ashort silica gel column with 20:1, 10:1, 6:1 Hex/EtOAc. It gave 11.6 gof the iodide 531-YW-3.

[0361] Coupling:

[0362] To a solution of iodide (531-YW-3, 11.6 g, 31.3 mmol) andselenide (509-HD-213, 24.5 g, 50.9 mmol, 1.6 eq.) in a mixed solvent ofTHF and HMPA (130 mL, 10:1 ratio), a solution of LiHMDS (94 mL, 0.5 M)was added by a syringe pump in one and half hour at −78° C. After 20 minat −78° C., it was warmed to 0° C. After cooled back to −10° C., it wasquenched with aq. sat. NH₄Cl and extracted with EtOAc (2×). The organiclayers were washed with brine, dried and concentrated. The crude productwas purified on silica gel with Hexanes/EtOAc, 20:1, 10:1, 6:1, 4:1, and2:1 to give 16.0 g.

[0363] The product from the above (16 g) was dissolved in CH₂Cl₂ (200mL), MCPBA (16 g, 50.9 mmol, 1.6 eq., 55%) was added at 0° C. After 15min at 0° C., Triethylamine (20 mL, excess) was added. After 30 min, itwas quenched with aq. Sat. Na₂S₂O₃. After stirred for 20 min, it wasextracted with EtOAc (2×). The organic layers were washed with sat.Na₂S₂O₃, sat. NaHCO₃, brine, dried and concentrated. The crude productwas purified on silica gel with Hexanes/EtOAc, 20:1, 10:1, 3:1, gave12.5 g of the desired product, 531-YW-4 (83% in three steps).

[0364] To a solution of ester (5.66 g, 10 mmol) In EtOH (100 mL), 50 mLof 1N NaOH was added. The reaction was stirred at rt overnight. Thereaction was then diluted with water, extracted with EtOAc (3×). Thecombined organic layers were washed with brine, dried and concentrated.It was purified on silica gel column to give 4.42 g of desired productas an oil (92%).

[0365] To a solution of (COCl)₂ (2.2 mL, 3 eq.) in 50 mL of CH₂Cl₂, DMSOwas added slowly at −78° C. After 15 min at −78° C., a solution ofalcohol (4.1 g, 3.5 mmol) was added into the reaction at −78° C. After30 min at that temperature, TEA (10.7 mL, 9 eq.) was added. The reactionwas warmed to rt. It was quenched with Sat. NH₄Cl, extracted with EtOAc(2×). The combined organic layers were washed with brine, dried andconcentrated to dryness. It was used in next reaction withoutpurification.

[0366] The aldehyde was used as a general intemediate for the synthesisof C3-C4 modification by coupling with appropiate acetylene orequivalent.

[0367] Preparation of Acetylenes for C3-C4 Modifications:

[0368] To a solution of TMS-acetylene (38.8 mL) In 1 L of THF at −60°C., n-BuLi (110 mL, 2.5 M) was added. The reaction was warmed to 0° C.briefly, then cooled back down to −60° C. BF₃Et₂O (33.8 mL) was thenslowly added, followed by the epoxide (15 mL) via syringe pump. Afterstirred at −60° C. for 1.5 h, it was warmed to rt, quenched by Sat.NH₄Cl, extracted with EtOAc (2×). The organic layers were dried andconcentrated. The crude product was purified by silica gel column with4:1 Hexanes/EtOAc to give 13.9 g of desired product as an oil.

[0369] The alcohol was silylated under standard condition with TBSCl andImidazole in methylene chloride.

[0370] A mixture of TBS protected TMS-acetylene (8.7 g) and K₂CO₃ (8 g)in methanol (120 mL) were stirred for 5 h at rt. It was extracted withEtOAc (2×). The organic layers were dried concentrated. The crudeproduct was purified on silica gel with hexanes to give 6.17 g ofcolorless oil (95%)

[0371] The following acetylenes were prepared analogous to thepreparation described:

[0372] Preparation of ER803064:

[0373] Acetylene (2.65 g, 12.5 mmol) was dissolved in 30 mL of THF andcooled down to −78° C. n-BuLi (2.5 N, 6.24 mL) in Hexane was added. Thereaction mixture was stirred at −78° C. for 10 min, and then a solutionof aldehyde (3.0 g, 6.24 mmol) in 30 mL of THF was added via cannula.The reaction mixture was stirred at −78° C. for 30 min and warmed upgradually to room temperature. It was quenched with water and extractedwith EtOAc. After purification on silica gel column, 3.7 g of 509-HD-108was obtained as a pale yellow oil in 78% yield.

[0374] 509-HD-108 (3.4 g, 4.91 mmol) was dissolved in 200 mL hexane.Quinoline (200 μL) and Lindlar catalyst (500 mg) were added. Thereaction mixture was stirred at 40° C. under H₂ balloon atmosphere for atotal of 18 h. Then the catalyst was filtered away. Quantitative amountof 509-HD-112 was obtained as a pale yellow oil.

[0375] 509-HD-112 (3.4 g, 4.9 mmol) was dissolved in 60 mL ofdichloromethane at room temperature. Triethylamine (1.71 mL, 9.8 mmol),benzoyl chloride (1.14 mL, 12.2 mmol) and catalytic amount of DAMP wereadded, respectively. After stirring for 12 h, 0.1N sodium hydroxidesolution was added and the reaction mixture was extracted with EtOAc.The crude product was purified on silica gel column, giving 509-HD-115as a colorless oil in 94% yield.

[0376] 509-HD-115 (3.7 g, 4.64 mmol) was dissolved in 50 mL of THF. TheTHF solution of TBAF (1N, 25 mL) was added. The reaction mixture washeated at 50° C. for 24 h. It was diluted with Et₂O and washed with H₂O.After purification on silica gel column, 509-HD-116 was obtained as apale yellow foam in 68% yield.

[0377] 2-Chloro-1-methylpyridinium iodide (2.4 g, 9.5 mmol) and n-Bu₃N(2.3 mL, 9.5 mmol) were dissolved in 180 mL of dichloromethane andheated to reflux. The solution of 509-HD-116 (1.85 g, 3.2 mmol) in 50 mLof THF was added slowly. The reaction mixture was heated for 30 min. Itwas washed with 0.02 N hydrochloric acid, sat. sodium bicarbonatesolution and brine, respectively. After purification on silica gelcolumn, 509-HD-118 was obtained as a pale yellow foam in 62% yield.

[0378] 509-HD-118 (1.22 g, 2.2 mol) was dissolved in 30 mL of ethanol.Sodium hydroxide (1N, 21.5 mL) solution was added. The reaction mixturewas stirred for 48 h at room temperature. It was diluted with H₂O,extracted with EtOAc. After purification on silica gel column, 346 mg ofthe major desired single isomer 509-HD-119B was obtained as a colorlessoil.

[0379] 509-HD-119B (155 mg, 0.34 mmol) was dissolved in 9 mL ofdichloromethane. Molecular sieve (4A, 360 mg) and PCC (360 mg, 1.7 mmol)were added. The reaction mixture was stirred for 1 h at roomtemperature. After passing through celite, 509-HD-125 was obtained ascolorless oil in quantitative yield.

[0380] 509-HD-125 was dissolved in 2.5 mL of dichloromethane. Thenhydrofluoric acid (6 N, 10 mL) was added. The reaction mixture wasstirred at room temperature for 30 min. It was diluted with moredichloromethane, washed with water and sat. sodium bicarbonate solution.After purification on a plug of silica gel, ER803064 was obtained as awhite solid in 86% yield.

[0381] ER805149 were synthesized in similar manner starting from thecorresponding acetylene.

[0382] Preparation of B2526 (Trans Cyclopentane with Desired DiolConfiguration)

[0383] Step 1

[0384] To a solution of (trimethylsilyl) acetylene (8.8 mL, 62 mmol) indry THF (100 mL), at −78° C. under an inert atmosphere, were added a2.5M solution of n-butyllithium (24.9 mL, 62 mmol) and boron trifluoridediethyl etherate (7.66 mL, 62 mmol). The reaction mixture was thentreated drop wise with a solution of cyclopentene oxide (2.71 mL, 31mmol). The reaction mixture was stirred at −78° C. for 1 hour thenwarmed to room temperature. The usual work up gave compound 453-MS-226(3.92 g; 69%).

[0385] Step 2

[0386] To a mixed solution of 453-MS-226 (3.77 g, 20.67 mmol) and4-methoxybenzyl 2,2,2-trichloroacetimidate (7 g, 24.8 mmol) in diethylether (80 mL), at room temperature under an inert atmosphere, was addeda 1M solution of trifluoromethane sulphonic acid in diethyl ether (0.62mL) drop wise over approximately 15 minutes. Two extra aliquots (1 mLeach) of the 1M trifluoromethane sulphonic acid solution were added at10 minutes and at 60 minutes. The usual work up, with subsequent partialpurification by chromatography, gave impure compound 453-MS-228 (3.37 g,approximately 54%), which was used in the next step without furtherpurification.

[0387] Step 3

[0388] A solution of impure 435-MS-228 (3.37 g, approximately 0.011 mol)in methanol (33 mL) was treated with potassium carbonate (3.075 g, 0.022mol) and stirred for 3.5 hours. The usual work up, followed bychromatographic purification, gave compound 453-MS-230 (2.22 g,approximately 88%).

[0389] Step 4

[0390] To a solution of 453-MS-230 (1 g, 4.37 mmol) in dry THF (10 mL),at −78° C. under an inert atmosphere, was added drop wise a 1.6Msolution of n-butyllithium (2.73 mL, 4.37 mmol). The reaction mixturewas stirred at −78° C. for 10 minutes, then warmed momentarily to 0° C.,and cooled to −78° C. A solution of the 343-YW-277 (1.5 g, 3.63 mmol) indry THF (15 mL) was added drop wise. The reaction mixture was stirred at−78° C. for 30 minutes then warmed to room temperature. The usual workup, followed by chromatographic purification, gave compound 453-MS-232(1.90 g, 82%).

[0391] Step 5

[0392] A solution of compound 453-MS-232 (1.68 g, 2.61 mmol) in hexane(30 mL) was hydrogenated, at room temperature and pressure, in thepresence of Lindlar's catalyst (168 mg) and quinoline (30 μL) for 20hours. Filtration and concentration in vacuo gave compound 453-MS-237(1.68 g, assumed quantitative) which was used in the next stage withoutpurification.

[0393] Step 6

[0394] To a solution of compound 453-MS-237 (1.61 g, 2.5 mmol) in drydichloromethane (20 mL) were added triethylamine (2.09 mL, 15 mmol),DMAP (30 mg, 0.25 mmol), and benzoyl chloride (0.58 mL, 5 mmol). Thereaction mixture was stirred at room temperature for 3 days then workedup in the usual manner. Chromatographic purification gave compound453-MS-240 (1.29 g, 69%).

[0395] Step 7

[0396] To a solution of compound 453-MS-240 (300 mg, 0.4 mmol) in THF (6mL) was added TBAF (210 mg, 0.8 mmol). The reaction mixture was stirredat room temperature for 2.5 hours then worked up in the usual manner togive compound 453-MS-244 (145 mg, 71%) (m/z: 533.2516 measured [M+23],533.2510 calculated).

[0397] Step 8

[0398] A mixed solution of compound 453-MS-244 (737 mg, 1.44 mmol) andtriphenylphosphine (682 mg, 2.6 mmol) in dry toluene (10 mL) was cooledto 0° C. under an inert atmosphere. A solution of dibenzylazidodicarboxylate (1.033 g, 3.46 mmol) in toluene (5 mL), and methyliodide (117 μL; 1.88 mmol) were added, separately and simultaneously, tothe reaction mixture over approximately 15 seconds. The reaction mixturewas stirred at 0° C. for 15 minutes then allowed to warm to roomtemperature. After 30 minutes at room temperature the reaction mixturewas worked up in the usual manner. Chromatographic purification gavecompound 453-MS-253 (550 mg, 61%).

[0399] Step 9

[0400] A mixture of compound 453-MS-253 (475 mg, 0.765 mmol) andcompound 554-RB-260 (519 mg, 0.86 mmol) was dissolved in a solution of10% HMPA in THF (6 mL) and cooled to −78° C. under an inert atmosphere.A 0.5 M solution of LiHMDS in THF (1.83 mL, 0.916 mmol) was then addeddrop wise over approximately 30 minutes. The reaction mixture wasstirred at −78° C. for 1 hour then treated with a 1M solution of LiHMDSin THF (0.916 mL; 0.916 mmol). After 20 minutes the reaction mixture waswarmed to 0° C. The intermediate crude product was worked up in theusual manner and purified partially by chromatography. The intermediatewas dissolved in dichloromethane (8 mL) and cooled to 0° C. A solutionof approximately 65% meta-chloroperbenzoic acid (249 mg) indichloromethane (2 mL) was added portion wise. After 30 minutestriethylamine (0.65 mL) was added and the usual work up, followed bychromatographic purification, gave compound 453-MS-262 (348 mg, 47%).

[0401] Step 10

[0402] To a vigorously stirred biphasic mixture of compound 453-MS-262(440 mg, 0.538 mmol), dichloromethane (20 mL) and water (10 mL), wasadded drop wise a solution of DDQ (47 mg, 0.646 mmol) in dichloromethane(15 mL). After 1 hour at room temperature the reaction mixture wasworked up in the usual manner. Chromatographic purification gave twofractions of partially resolved diastereoisomers: Fraction A (lesspolar): a mixture of 3 co-eluted diastereoisomers—compound 453-MS-277A(190 mg); Fraction B (more polar): a single diastereoisomer—compound453-MS-277B (122 mg); (total yield: 312 mg, 83%)

[0403] Step 11

[0404] A solution of compound 453-MS-277B (122 mg, 0.175 mmol) intetrahydrofuran (5 mL) was treated with a solution of TBAF (92 mg, 0.35mmol) in THF (1 mL). The reaction mixture was stirred at roomtemperature for 6 hours then worked up in the usual manner to giveimpure compound 453-MS-279 (104 mg), which was used in the next stagewithout purification.

[0405] Step 12

[0406] A solution of crude compound 453-MS-279 (80 mg, assumed tocontain 0.134 mmol) in dichloromethane (30 mL) was treated with2-chloro-1-methylpyridinium iodide (45 mg, 74 mmol) and tri-n-butylamine(42 μL, 0.174 mmol). The reaction mixture was heated under reflux for 25minutes then cooled to room temperature. The usual work up andchromatographic purification gave compound 453-MS-284 (30 mg, 39% fromcompound 453-MS-277B).

[0407] Step 13

[0408] A solution of compound 453-MS-284 (30 mg, 51 μmol) in a mixtureof ethanol (1 mL) and tetrahydrofuran (0.5 mL) was treated with 1Maqueous sodium hydroxide (518 μL) and stirred for approximately 3 daysat room temperature. The usual work up, followed by chromatographicpurification, gave compound 453-MS-289 (16 mg, 65%).

[0409] Step 14

[0410] A solution of compound 453-MS-289 (15 mg, 31.6 μmol) indichloromethane (1.5 mL) was treated with PCC (81 mg, 0.375 mmol) in thepresence of powdered 4 Å molecular sieves (81 mg). The reaction mixturewas stirred vigorously for 70 minutes at room temperature. Basificationwith excess triethylamine, followed by partial chromatographicpurification gave impure compound 453-MS-296 (approximately 7 mg), whichwas used in the next stage without further purification.

[0411] Step 15

[0412] A solution of impure compound 453-MS-296 (approximately 7 mg) ina mixture of acetonitrile (1600 μL) and dichloromethane (400 μL) wastreated with 48% aqueous hydrofluoric acid (400 μL). After 25 minutesthe usual work up followed by chromatographic purification gave compoundB2526 (1.1 mg, approximately 6% from compound 453-MS-289).

[0413] Preparation of B2538 (Cis Cyclopentane with Desired DiolConfiguration)

[0414] Step 1

[0415] To a solution of compound 453-MS-277A (190 mg, 0.273 mmol) intetrahydrofuran (7 ml:) was added a solution of TBAF (143 mg, 0.545mmol) in THF (2 mL). After 1 hour at room temperature the reactionmixture was treated with additional TBAF (20 mg, 0.076 mmol). After afurther 3 hours the reaction mixture was worked up in the usual mannerto give impure compound 453-MS-281 (186 mg), which was used in the nextstage without purification.

[0416] Step 2

[0417] To a solution of triphenylphosphine (31.5 mg, 0.12 mmol) in drytetrahydrofuran (2.5 mL) was added diethyl azidodicarboxylate (19 mL;0.12 μmol), at room temperature under an inert atmosphere. A solution ofimpure compound 453-MS-281 (36 mg, 0.06 mmol) in dry tetrahydrofuran(2.5 mL) was added. After 90 minutes additional triphenylphosphine (31.5mg, 0.12 mmol) and diethyl azidodicarboxylate (19 ml, 0.12 μmol) wereadded. After a further 30 minutes the reaction mixture was worked up inthe usual manner. Chromatographic purification gave compound 501-MS-6(19 mg, 54% from compound 453-MS-277A).

[0418] Step 3

[0419] A solution of compound 501-MS-6 (19 mg, 32.8 μmol) in a mixtureof ethanol (1 mL) and THF (0.5 mL) was treated with 1M aqueous NaOH (380μL) and stirred for approximately 17 hours at room temperature. Thereaction mixture was then heated to 100° C. for approximately 30minutes. The usual work up gave compound 501-MS-8 (15.5 mg,quantitative).

[0420] Step 4

[0421] A solution of compound 501-MS-8 (15.5 mg, 32 μL) indichloromethane (3.2 treated with PCC (85 mg, 0.39 mmol) in the presenceof powdered 4 Å molecular sieves (85 mg). The reaction mixture wasstirred vigorously for 2 hours at room temperature. Basification withexcess triethylamine, followed by chromatographic purification gavecompound 501-MS-11 (12.5 mg, 83%).

[0422] Step 5

[0423] A solution of compound 501-MS-11(12 mg, 25 μmol) in a mixture ofacetonitrile (2400 μL) and dichloromethane (600 μL) was treated with 48%aqueous hydrofluoric acid (600 μL). After 1 hour the usual work upfollowed by chromatographic purification gave compound B2538 (4 mg, 41%)(m/z: 411.1 [M+23, 100%], 412.1 [35%]).

[0424] Preparation of B2522 (Trans Cyclopentane with Undesired DiolConfiguration)

[0425] Step 1

[0426] A solution of compound 453-MS-277A (190 mg, 0.273 mmol) in THF (7mL) was treated with a solution of TBAF (143 mg, 0.545 mmol) in THF (2mL). The reaction mixture was stirred at room temperature for 6 hoursthen worked up in the usual manner to give impure compound 453-MS-281(186 mg), which was used in the next stage without purification.

[0427] Step 2

[0428] A solution of crude compound 453-MS-281 (150 mg, assumed tocontain 0.251 mmol) in dichloromethane (15 mL) was treated with2-chloro-1-methylpyridinium iodide (84 mg, 0.327 mmol) andtri-n-butylamine (78 μL, 0.327 mmol). The reaction mixture was heatedunder reflux for 40 minutes then treated with additional2-chloro-1-methylpyridinium iodide (84 mg, 0.327 mmol) andtri-n-butylamine (78 μL, 0.327 mmol). The reaction mixture was heatedunder reflux for a further 1 hour then cooled to room temperature. Theusual work up and chromatographic purification gave compound 453-MS-290(72 mg, 50% from compound 453-MS-277A).

[0429] Step 3

[0430] A solution of compound 453-MS-290 (72 mg, 0.124 mol) in a mixtureof ethanol (2.4 mL) and THF (1.2 mL) was treated with 1M aqueous NaOH(1.24 mL, 1.24 mmol) and stirred for approximately 4 days at roomtemperature. The usual work up, followed by chromatographicpurification, gave three fractions of partially resolved compounds:

[0431] Fraction A (less polar): an unascertained mixture ofdiastereoisomers (10 mg);

[0432] Fraction B (more polar): a mixture of 2 diastereoisomers—compound453-MS-292B (46 mg);

[0433] Fraction C (most polar): single diastereoisomer of startingmaterial 453-MS-290 (2 mg);

[0434] (total yield: 56 mg; 95%)

[0435] Step 4

[0436] A solution of compound 453-MS-292B (20 mg, 42 μmol) indichloromethane (2 mL) was treated with PCC (109 mg, 0.505 mmol) in thepresence of powdered 4 Å molecular sieves (109 mg). The reaction mixturewas stirred vigorously for 55 minutes at room temperature. Basificationwith excess triethylamine, followed by chromatographic purification,gave compound 453-MS-299 (17 mg, 86%).

[0437] Step 5

[0438] A solution of compound 453-MS-299 (19 mg, 0.04 mmol) in a mixtureof acetonitrile (4 mL) and dichloromethane (1 mL) was treated with 48%aqueous hydrofluoric acid (1 mL). After 35 minutes the usual work upfollowed by chromatographic purification gave compound B2522 (9 mg, 58%)(m/z: 411.1443 measured [M+23], 411.1420 calculated).

[0439] ER804018 and ER804019 (C4-F):

[0440] LDA (2.0 M , 36 mL) was added to the solution of ethyl2-fluoropropanate (7.23 g) and acetaldehyde (13.5 mL) in ether (100 mL)at −78° C. After the addition finished the reaction flask was kept inice bath and gradually warm to room temperature. The reaction wasquenched with aqueous ammonium chloride after overnight stirring. Theaqueous phase was extracted with ether and the combined organic phasewas dried over sodium sulfate. The solvent was stripped off and theresidue was distilled in vacuo to give 541-YJ-97 (4.22 g).

[0441] Chloro-t-butyldiphenylsilane was added to the mixture of541-YJ-97 (4.22 g) and imidazole (3.5 g) in methylene chloride (50 mL)and stirred overnight. Aqueous sodium bicarbonate was added to thereaction mixture. The aqueous phase was extracted with ether and thecombined organic phase was dried over sodium sulfate. The solvent wasstripped off and the residue was purified with flush chromatograph(hexane/acetate 50/1) to give 541-YJ-99 (7.55 g).

[0442] DIBAL-H in toluene (1.0 M , 25 ml) was added to 541-YJ-99 intoluene (85 mL) at 0° C. The reaction was quenched in one hour withmethanol/water and filtrated through Celite. The residue was purified byflash chromatograph to yield 541-YJ-101 (536 mg).

[0443] Dess-Martin periodinane was added to 541-YJ-101 (511 mg) inmethylene chloride (15 ml) at room temperature. The mixture was dilutedwith ether in 40 minutes and filtrated through Celite. The filtrate wasconcentrated, and the residue was purified by prepTLC (hex/acetate 7/1)to give 541-Yj-105 (355 mg, 70%).

[0444] Triphenylphosphine (2.08 g) was added to carbon tetrabromide(1.31 g) in methylene chloride at room temperature. After stirring for40 minutes 541-YJ-105 was added and stirred for 2 hours. The mixture wasconcentrated and filtrated through silica gel (hexane/acetate 7/1) toproduce 541-YJ-106 (452 mg, 89%).

[0445] n-Butyllithium (2.5 M , 0.74 mL) was added to 541-YJ-106 (450 mg)in THF (10 mL) at −78° C. After one hour 541-YJ-108 was added. Thereaction was kept at 0° C. for one hour and warmed to room temperaturebefore it was quenched with aqueous ammonium chloride. The aqueous phasewas extracted with ether and the combined organic phase was dried oversodium sulfate. The solvent was stripped off and the residue waspurified with TLC (hexane/acetate 4/1) to 541-YJ-109 (394 mg, 54%).

[0446] The suspension of Lindlar catalyst (420 mg) and 541-YJ-109 (390mg) in hexane (8 mL) was charged with hydrogen and stirred overnight.The suspension was filtrated through Celite and rinsed with acetate. Thefiltrate was concentrated to give 541-YJ-111 (378 mg).

[0447] Benzoyl chloride (254 mg) and DMAP (catalytic amount) was addedto the solution of 541-YJ-111 (378 mg) and triethylamine (0.5 mL) inmethylene chloride (7 mL) at room temperature. The mixture was keptstirring overnight and quenched with aqueous sodium bicarbonate. Theaqueous phase was extracted with ether and the combined organic phasewas dried over sodium sulfate. The solvent was stripped off and theresidue was purified with TLC (hexane/acetate 7/1) to 541-YJ-115 (419mg).

[0448] The solution of 541-YJ-115 (419 mg) and TBAF (1.0 M , 2.2 mL) inTHF (8 mL) was kept stirring overnight, and then diluted with water. Theaqueous phase was extracted with ether and concentrated. The residue waspurified with TLC (methylene chloride/methanol 10/1) to give 541-YJ-116(194 mg).

[0449] The solution of 541-YJ-115 (419 mg) and TBAF (1.0 M , 2.2 mL) inTHF (8 mL) was kept stirring overnight, and then diluted with water. Theaqueous phase was extracted with ether and concentrated. The residue waspurified with TLC (methylene chloride/methanol 10/1) to give 541-YJ-116′(21 mg).

[0450] 541-YJ-116 (21 mg) was added to the reflux of2-chloro-1-methylpyridium iodide (32 mg) and tributylamine (23 mg) inmethylene chloride (4 mL). After 2 hours reflux the mixture was stirredovernight. The mixture was diluted with ether and washed with HCl (1.0N) and water. The residue was purified with TLC (hexane/acetate 1/1) togive 541-YJ-118-1 (7.7 mg) and 541-YJ-118-2 (8.4 mg).

[0451] PCC was added to 541-YJ-118-1 (7.7 mg) and MS 4A suspension inmethylene chloride (2 mL) at room temperature. The mixture was stirredfor 3 hours and filtrated through silica gel. The silica gel was elutedwith acetate and concentrated to give 541-YJ-119 (3.3 mg).

[0452] PCC was added to 541-YJ-118-2 (8.4 mg) and MS 4A suspension inmethylene chloride (2 mL) at room temperature. The mixture was stirredfor 3 hours and filtrated through silica gel. The silica gel was elutedwith acetate and concentrated to give 541-YJ-120 (3.0 mg).

[0453] Hydrofluoric acid (49%, 1 mL) was added to 541-YJ-119 (8.0 mg) inacetonitrile (3 mL) and stirred for 15 minutes. The mixture was dilutedwith water and extracted with methylene chloride. The organic phase wasconcentrated and purified with a short silica gel pad to produce541-YJ-126 (5.1 mg, ER-804018).

[0454] Hydrofluoric acid (49%, 1 mL) was added to 541-YJ-120 (6.0 mg) inacetonitrile (3 mL) and stirred for 15 minutes. The mixture was dilutedwith water and extracted with methylene chloride. The organic phase wasconcentrated and purified with a short silica gel pad to produce541-YJ-126 (3.1 mg, ER-804019).

[0455] Preparation of ER804142 and ER804143, C4-CF3 Analog:

[0456] To neat trifluoropropanic acid (10.0 g) was added oxalyl chloride(7.5 mL) at room temperature and kept the mixture at 50° C. overnight.Then benzyl alcohol was added and kept stirring for 10 hours. Thereaction was quenched with aqueous sodium bicarbonate and extracted withchloroform. The solvent was stripped off and the residual was purifiedwith flush chromatograph (hexane/acetate 20/1) to afford 541-YJ-139(16.05 g, 94%).

[0457] LDA (1.5 M , 53.9 ml) was added to the solution of 541-YJ-139(7.23 g) and acetaldehyde (20.6 ml) in THF (180 mL) at −78° C. After theaddition finished the reaction flask was kept in ice bath and graduallywarm to room temperature. The reaction was quenched with aqueousammonium chloride after overnight stirring. The aqueous phase wasextracted with ether and the combined organic phase was dried oversodium sulfate. The solvent was stripped off and the residue waspurified with flush chromatograph (hexane/acetate 20/1 to 10/1) toafford 541-YJ-141 (10.26 g, 53%).

[0458] Chloro-t-butyldiphenylsilane (12.2 ml) was added to the mixtureof 541-YJ-141 (10.26 g) and imidazole (10.7 g) in methylene chloride(200 ml) and stirred overnight. Aqueous sodium bicarbonate was added tothe reaction mixture. The aqueous phase was extracted with chloroformand the combined organic phase was dried over sodium sulfate. Thesolvent was stripped off and the residue was purified with flushchromatograph (hexane/acetate 40/1) to give 541-YJ-143 (14.94 g, 76%).

[0459] DIBAL-H in toluene (1.0 M , 89.4 mL) was added to 541-YJ-143 intoluene (300 mL) at 0° C. The reaction was quenched in one and half hourwith methanol/water (70 mL/45 mL) and filtrated through Celite. Theresidue was purified by flash chromatograph (hexane/acetate 10/1) toyield 541-YJ-144 (7.38 g, 62%).

[0460] Dess-Martin periodinane (9.47 g) was added to 541-YJ-144 (7.38 g)in methylene chloride (150 mL) at room temperature. The mixture wasdiluted with ether in one hour and filtrated through Celite. Thefiltrate was concentrated and the residue was purified by flashchromatograph (hexane/acetate 10/1) to yield 541-YJ-145 (7.37 g).

[0461] Triphenylphosphine (39.2 g) was added to carbon tetrabromide(24.74 g) in methylene chloride at room temperature. After stirring for45 minutes 541-YJ-145 (7.37 g) was added and stirred for 3 hours. Themixture was concentrated and purified by flash chromatograph (methylenechloride) to yield 541-YJ-146 (8.74 g, 85%).

[0462] n-Butyllithium (2.5 M , 3.47 mL) was added to 541-YJ-106 (2.39 g)in THF (20 mL) at −78° C. After one hour 343-YW-277 (0.98 g) was addedat −78° C. The reaction was kept at 0° C. for one hour and then warmedto room temperature before it was quenched with aqueous ammoniumchloride. The aqueous phase was extracted with ether and the combinedorganic phase was dried over sodium sulfate. The solvent was strippedoff and the residue was purified with flash chromatograph(hexane/acetate, 3/1) to 541-YJ-148 (1.40 g, 65%).

[0463] The suspension of Rieke zinc (9.0 mL) was carefully added to541-YJ-148 (390 mg) in methanol-water (20 mL/2 mL) at room temperature.The suspension was heated at 70° C. for one and half hour. The mixturewas filtrated through Celite and rinsed with acetate. The filtrate wasconcentrated to give 541-YJ-151 (1.09 g) without purification.

[0464] Benzoyl chloride (0.52 ml) and DMAP (catalytic amount) was addedto the solution of 541-YJ-151 (0.97 g) and triethylamine (1.24 mL) inmethylene chloride (15 mL) at room temperature. The mixture was keptstirring overnight and quenched with aqueous sodium bicarbonate. Theaqueous phase was extracted with chloroform and the combined organicphase was dried over sodium sulfate. The solvent was stripped off andthe residue was purified with flash chromatograph (hexane/acetate 5/1)to 541-YJ-158 (0.96 g, 88%).

[0465] The solution of 541-YJ-158 (0.96 g) and TBAF (1.0 M , 4.9 mL) inTHF (20 mL) was kept at 50° C. overnight and then diluted with water.The aqueous phase was extracted with ether and concentrated. The residuewas purified with flash chromatograph (acetate) to give 541-YJ-159 (186mg, 30%).

[0466] 541-YJ-159 (186 mg) was added to the reflux of2-chloro-1-methylpyridium iodide (223 mg) and tributylamine (162 mg) inmethylene chloride (25 mL). After 2 hours reflux the mixture was cooleddown. The mixture was diluted with ether and washed with HCl (1.0 N) andwater. The residue was purified with TLC (hexane/acetate 1/1) to give541-YJ-160 (169 mg).

[0467] The solution of 593-YJ-160 (169 mg) and sodium hydroxide (1.0 N,0.5 mL) in ethanol (5 mL) was kept at 75° C. overnight. The mixture wasconcentrated and diluted with aqueous ammonium chloride. The aqueousphase was extracted with ether and the combined organic phase wasconcentrated. The residue was purified by TLC (hexane/acetate, 1/1) toyield 593-YJ-161 (39 mg, 28%).

[0468] Dess-Martin periodinane (50 mg) was added to 541-YJ-161 (39 mg)in methylene chloride (2 mL) at room temperature. The mixture wasstirred for 3 hours and diluted with ether. The mixture was filtratedthrough Celite and purified with TLC (hexane/acetate, 2/1) to give541-YJ-168-1 (6.7 mg) and 541-YJ-168-2 (5.3 mg).

[0469] Hydrofluoric acid (49%, 1 mL) was added to 541-YJ-168-1 (6.7 mg)or 541-YJ-168-2 (5.3 mg) in acetonitrile (3 mL) and stirred for 15minutes. The mixture was diluted with water and extracted withchloroform. The organic phase was concentrated and purified with TLC(hexane/acetate 1/4) to produce 541-YJ-174 (1.5 mg, ER-804142) or541-YJ-175 (0.5 mg, ER-804143).

[0470] Preparation of C4-Oxo Analogs, NF0675, NF0879, NF0880, and NF0905

[0471] Synthetic Procedure for NF0675

[0472] Methyl S-lactate (20.8 g, 0.2 mol) was dissolved in dry THF (500mL), imidazole (17.7 g, 0.26 mol) was added and the mixture was cooledto 0° C. in ice/water bath. Then TBDPSCl (60.5 g, 0.22 mol) was added,the mixture was allowed to warm slowly to rt and stirred overnight afterwhich a saturated solution of NaHCO₃ was added. The mixture wasextracted with EtOAc and the organic extract was washed with a saturatedsolution of NaHCO₃, water, brine, dried with anhydrous Na₂SO₄, filteredand concentrated.

[0473] The crude product (74.59 g) was dissolved in dry Et₂O (300 mL)and the solution was cooled to 0° C. in ice/water bath. Then LiBH₄ (4.36g, 0.2 mol) was added portionwise, the mixture was allowed to warmslowly to rt and stirred for 2 days after which a saturated solution ofNH₄Cl was added slowly. The mixture was extracted with EtOAc and theorganic extract was washed with a saturated solution of NH₄Cl, water,brine, dried with anhydrous Na₂SO₄, filtered and concentrated. The crudeproduct was purified by chromatography on silica gel using 20%EtOAc/hexane to give 59.67 g (0.19 mol, 95% 2 steps) of the protectedcompound TM-01.

[0474] To a solution of oxalyl chloride (2.5 eq., 75 mmol, 6.54 mL) inCH₂Cl₂ (60 mL), DMSO (5 eq., 150 mmol, 10.6 mL) was added at −78° C.After 15 min at −78° C, a solution of alcohol TM-01 (30 mmol, 9.44 g) inCH₂Cl₂ (100 mL) was added over a period of 40 min. After 30 min at −78°C., Et₃N (6.5 eq., 195 mmol, 27.2 mL) was added and the reaction waswarmed to −50° C. and stirred for 30 min. It was quenched with asaturated solution of NH₄Cl (100 mL), extracted with EtOAc. The organiclayer was dried with Na₂SO₄, filtered and concentrated. The crudealdehyde (9.55 g) was dissolved in dry THF (100 mL) and cooled to −78°C. Then 0.5M solution of propargyl magnesium bromide in dry THF (1.7eq., 50 mmol, 100 mL) was added dropwise over a period of 30 min and thereaction was warmed to −10° C. It was quenched with a saturated solutionof NH₄Cl, extracted with EtOAc. The organic layer was dried with Na₂SO₄,filtered and concentrated. The crude alcohol was purified bychromatography on silica gel using 5% EtOAc/hexane to give 5.125 g (15.1mmol, 50% 2 steps) of the desired alcohol.

[0475] The alcohol (5.124 g, 15.1 mmol) was dissolved in CH₂Cl₂ (55 mL),diisopropylethylamine (15.84 mL, 90.8 mmol) was added and the mixturewas cooled to 0° C. in ice/water bath. Then chloromethyl methyl ether(3.45 mL, 45.4 mmol) was added and the mixture was allowed to warm tort. After 2 days, it was quenched with a saturated solution of NH₄Cl,extracted with EtOAc. The organic layer was dried with Na₂SO₄, filteredand concentrated. The crude product was purified by chromatography onsilica gel using 5% EtOAc/hexane to give 4.866 g (12.7 mmol, 84%) ofTM-02.

[0476] 491-HAD-46 (620 mg, 2.4 mmol) was dissolved in dry DMF (10 mL),imidazole (243 mg, 3.6 mmol) was added and the mixture was cooled to 0°C. in ice/water bath. Then TBSCl (430 mg, 2.9 mmol) was added, themixture was allowed to warm slowly to rt and stirred for 30 min afterwhich a saturated solution of NaHCO₃ was added. The mixture wasextracted with EtOAc and the organic extract was washed with a saturatedsolution of NaHCO₃, water, brine, dried with anhydrous Na₂SO₄, filteredand concentrated. The crude product was purified by chromatography onsilica gel using 5% EtOAc/hexane to give 856 mg (2.3 mmol, 96%) of thesilyl ether.

[0477] To a suspension of LiAlH₄ (65 mg, 1.7 mmol) in dry THF (7 mL) wasadded a solution of the silyl ether (856 mg, 2.3 mmol) in dry THF (13.5mL) at 0° C. The mixture was allowed to warm slowly to rt and stirredfor 50 min after which EtOAc and 1N HCl were added. The mixture wasextracted with EtOAc and the organic extract was washed with water, asaturated solution of NH₄Cl, water, brine, dried with anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 25% EtOAc/hexane to give 654 mg (2.3mmol, 99%) of the alcohol.

[0478] The alcohol (654 mg, 2.3 mmol) was dissolved in dry CH₂Cl₂ (25mL). Then Dess-Martin periodinane (1.67 g, 3.94 mmol) was added andstirred for 4 hr after which a saturated solution of Na₂S₂O₃ and asaturated solution of NaHCO₃ were added. The mixture was extracted withEtOAc and the organic extract was washed with a saturated solution ofNaHCO₃, water, brine, dried with anhydrous Na₂SO₄, filtered andconcentrated to give 648 mg (2.3 mmol, quant.) of TM-03.

[0479] TM-02 (2.2 eq., 5.0 mmol, 1.89 g) was dissolved in THF (20 mL)and cooled to −78° C., under nitrogen. Then, n-BuLi (1.6M/hexane, 2.0eq., 4.5 mmol, 2.8 mL) was added and the reaction was stirred at −78° C.for 60 min. Aldehyde TM-03 (2.3 mmol, 648 mg) dissolved in THF (8 mL)was added to the solution and stirred for 60 min at −78° C. The solutionwas allowed to warm to rt and stirred for 1.5 hrs. The mixture wasquenched with water, extracted with EtOAc and the organic extract waswashed with brine, dried with Na₂SO₄, filtered and concentrated. Theresidue was purified by chromatography on silica gel using 15%EtOAc/hexane to give 1.393 g (2.1 mmol, 92%) of TM-04.

[0480] TM-04 (2.1 mmol, 1.39 g) was dissolved in hexane (40 mL). Then,quinoline (27 mg) and 5% Pd—BaSO₄ on carbon (88 mg) were added. H₂balloon was mounted and the mixture was purged 5× with H₂. Reaction wasstirred under hydrogen. After 27 hrs, reaction was stopped, catalyst wasfiltered through celite and mixture was concentrated under reducedpressure. The crude product was purified by chromatography on silica gelusing 15% EtOAc/hexane to give 957 mg (1.4 mmol, 69%) of TM-05 as majorisomer and 267 mg (0.4 mmol, 19%) of the diastereomer of allylic hydroxyposition.

[0481] Using same procedure for 554-RB-242, TM-05 (954 mg, 1.4 mmol) wasconverted to TM-06 (913 mg, 1.2 mmol, 83%).

[0482] TM-06 (912 mg, 1.2 mmol) was dissolved in THF (23 mL). Then,acetic acid (0.084 mL, 1.5 mmol) and 1.0M solution of tetrabutylammoniumfluoride in THF (1.23 mL, 1.23 mmol) were added at rt. The mixture wasstirred overnight after which a saturated solution of NH₄Cl was added.The mixture was extracted with EtOAc and the organic extract was washedwith a saturated solution of NaHCO₃, water, brine, dried with anhydrousNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel using 30% EtOAc/hexane to EtOAc as eluentsto give 362 mg (0.55 mmol, 47%) of TM-07 and 212 mg (0.50 mmol, 43%) ofTM-08.

[0483] Using same procedure for 554-RB-260, TM-07 (359 mg, 0.54 mmol)was converted to TM-09 (374 mg, 0.48 mmol, 89%).

[0484] Using same procedure for 5, TM-09 (372 mg, 0.48 mmol) wasconverted to TM-10 (339 mg, 0.35 mmol, 72%).

[0485] To a stirred solution of TM-10 (172 mg, 0.18 mmol) in THF (2 mL)and EtOH (2 mL) was added 1N NaOH aq. (2 mL) at rt. After 2.5 hrs, itwas quenched by 1N HCl and extracted with EtOAc. The organic extract waswashed with water, brine, dried with anhydrous Na₂SO₄, filtered andconcentrated. The crude product was purified by chromatography on silicagel using 30% EtOAc/hexane to give 143 mg (0.16 mmol, 93%) of the allylalcohol.

[0486] The allyl alcohol (143 mg, 0.16 mmol) was dissolved in THF (3mL). Then, 1.0M solution of tetrabutylammonium fluoride in THF (0.49 mL,0.49 mmol) was added at rt. The mixture was stirred for 3 hrs afterwhich 1N HCl was added. The mixture was extracted with EtOAc and theorganic extract was washed with water, brine, dried with anhydrousNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel using 10% MeOH/EtOAc to give 87 mg (0.16mmol, quant.) of TM-11.

[0487] To a stirred solution of TM-11 (87 mg, 0.16 mmol) in THF (3 mL)were added triethylamine (0.029 mL, 0.2 mmol) and 2,4,6-trichlorobenzoylchloride (0.032 mL, 0.2 mmol) at rt. After 16 hrs, the reaction mixturewas diluted with toluene (80 mL) and added dropwise to a solution ofN,N-dimethylaminopyridine (498 mg, 4.1 mmol) in toluene (80 mL) over aperiod of 8 hrs under reflux. The resultant mixture was stirred for 15hrs under reflux. After concentration under reduced pressure, theresidue was dissolved in EtOAc and washed with 5% citric acid aq.,water, brine and dried over anhydrous Na₂SO₄, filtered and concentrated.The crude product was purified by chromatography on silica gel using 30%EtOAc/hexane to give 53 mg (0.10 mmol, 64%) of TM-12.

[0488] Using similar procedure for 509-HD-125, TM-12 (39.4 mg, 0.078mmol) was converted to TM-13 (36.8 mg, 0.073 mmol, 94%).

[0489] To a stirred solution of TM-13 (12 mg, 0.024 mmol) in THF (1mL)-H₂O (0.5 mL) was added trifluoroacetic acid (1 mL) at 0° C. Themixture was then allowed to warm to rt. After 3.5 hrs, the mixture waspoured into a saturated solution of NaHCO₃ and extracted with EtOAc. Theorganic extract was washed with water, brine and dried over anhydrousNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel using 30% EtOAc/hexane to give 1.2 mg(0.0028 mmol, 12%) of NF0675.

[0490] Synthetic Procedure for NF0879 and NF0880

[0491] Using modified procedure for TM-02, TM-15 (10.56 g, 31.6 mmol)was obtained from 10.4 g (0.1 mol) of methyl S-lactate.

[0492] Using same procedure for TM-04, TM-03 (3.64 g, 12 mmol) wasconverted to TM-16 (5.29 g, 8.5 mmol, 69%).

[0493] Using same procedure for TM-05, TM-16 (5.28 g, 8.5 mmol) wasconverted to TM-17 (4.91 g, 7.9 mmol, 93%).

[0494] From 4.904 g (7.8 mmol) of TM-17, TM-19 (1.120 g, 1.8 mmol, 23%2steps) and TM-20 (2.218 g, 4.4 mmol, 57% 2steps) were obtained bysimilar procedure for TM-07.

[0495] Using similar procedure for TM-10, 1.104 g (1.8 mmol) of TM-19was converted to TM-22 (955 mg, 1.03 mmol, 58% 4steps).

[0496] Using similar procedure for TM-11, 955 mg (1.03 mmol) of TM-22was converted to TM-23 (593 mg, 0.98 mmol, 95% 2steps).

[0497] Using similar procedure for TM-12, 590 mg (0.98 mmol) of TM-23was converted to TM-24 (438 mg, 0.75 mmol, 77%).

[0498] Using similar procedure for TM-13, 209 mg (0.36 mmol) of TM-24was converted to TM-25 (186 mg, 0.32 mmol, 89%).

[0499] Using similar procedure for NF0675, 186 mg (0.32 mmol) of TM-25was ed to NF0879 (72 mg, 0.14 mmol, 45%).

[0500] NF0879 (16 mg, 0.032 mmol) was dissolved in CH₂Cl₂ (3 mL), H₂O(0.3 mL) and DDQ (2 eq., 0.064 mmol, 14.9 mg) were added and the mixturewas stirred vigorously at rt for 3 hrs. The mixture was quenched with asaturated solution of NaHCO₃ and diluted with EtOAc. The organic layerwas separated and washed with a saturated solution of NaHCO₃, brine,dried with Na₂SO₄, filtered and concentrated. The crude residue waspurified by chromatography on silica gel using 5% MeOH/CHCl₃ to give 5mg (0.013 mmol, 41%) of NF0880.

[0501] Synthetic Procedure for NF0905

[0502] TM-20 (2.218 g, 4.4 mmol) was dissolved in CH₂Cl₂ (45 mL),imidazole (520 mg, 7.6 mmol) was added and the mixture was cooled to 0°C. in ice/water bath. Then TBSCl (768 mg, 5.1 mmol) was added, themixture was allowed to warm slowly to rt and stirred for 1.5 hrs afterwhich a saturated solution of NaHCO₃ was added. The mixture wasextracted with EtOAc and the organic extract was washed with a saturatedsolution of NaHCO₃, water, brine, dried with anhydrous Na₂SO₄, filteredand concentrated to give 2.79 g of TM-26 as crude product.

[0503] TM-26 (2.79 g) was dissolved in CH₂Cl₂ (45 mL), triethylamine(1.85 mL, 13.3 mmol) and N,N-dimethylaminopyridine (54 mg, 0.44 mmol)were added and the mixture was cooled to 0° C. in ice/water bath. Thenbenzoyl chloride (1.03 mL, 8.9 mmol) was added, the mixture was allowedto warm slowly to rt and stirred overnight after which a saturatedsolution of NaHCO₃ was added. The mixture was extracted with EtOAc andthe organic extract was washed with a saturated solution of NH₄Cl, 5%citric acid aq., water, and brine, dried with anhydrous Na₂SO₄, filteredand concentrated to give 4.33 g of TM-27 as crude product.

[0504] TM-27 (4.33 g) was dissolved in CH₂Cl₂ (50 mL), H₂O (5 mL) andDDQ (1.28 g, 5.5 mmol) were added and the mixture was stirred vigorouslyat rt for 2 hrs. The mixture was quenched with a saturated solution ofNaHCO₃ and diluted with EtOAc. The organic layer was separated andwashed with a saturated solution of NaHCO₃ brine, dried with Na₂SO₄,filtered and concentrated. The crude residue was purified bychromatography on silica gel using 20% EtOAc/hexane to give 2.99 g ofTM-28 with slightly amount of impurities.

[0505] TM-28 (2.99 g) was dissolved in CH₂Cl₂ (60 mL),2,6-di-tert-butyl-4-methyl-pyridine (3.34 g, 16 mmol) and methyltrifrate (1.5 mL, 13 mmol) were added and the mixture was stirred underreflux overnight. The mixture was quenched with a saturated solution ofNaHCO₃ and diluted with EtOAc. The organic layer was separated andwashed with a saturated solution of NaHCO₃, brine, dried with Na₂SO₄,filtered and concentrated.

[0506] The crude residue (6.25 g) was dissolved in THF (60 mL). Then,1.0M solution of tetrabutylammonium fluoride in THF (5.5 mL, 5.5 mmol)was added at rt. The mixture was stirred for 1.5 hrs after which asaturated solution of NH₄Cl was added. The mixture was extracted withEtOAc and the organic extract was washed with water, brine, dried withanhydrous Na₂SO₄, filtered and concentrated. The crude product waspurified by chromatography on silica gel using 30% EtOAc/hexane to give362 mg (0.73 mmol, 16% 5 steps) of TM-29.

[0507] Using similar procedure for TM-09, 359 mg (0.72 mmol) of TM-29was converted to TM-30 (376 mg, 0.62 mmol, 86%).

[0508] Using similar procedure for TM-23, 371 mg (0.61 mmol) of TM-30was converted to TM-31 (207 mg, 0.42 mmol, 69% 5 steps).

[0509] Using similar procedure for TM-12, 207 mg (0.42 mmol) of TM-31was converted to TM-32 (206 mg, 0.42 mmol, quant.).

[0510] Using similar procedure for TM-13, 206 mg (0.42 mmol) of TM-32was converted to TM-33 (170 mg, 0.36 mmol, 83%).

[0511] Using similar procedure for NF0675, 170 mg (0.36 mmol) of TM-33was converted to NF0905 (50 mg, 0.13 mmol, 35%).

[0512] Preparation of Compound ER-804003 (C3 Trifluoromethyl)

[0513] Step 1

[0514] A solution of ethyl (R)-3-hydroxy-4,4,4-trifluorobutanoate (2 g,10.7 mmol) in DMF (20 mL) was treated with tert-butyldiphenylsilylchloride (8.34 mL, 32.1 mmol) and imidazole (3.28 g, 35.3 mmol). Thereaction mixture was heated at 70° C. under an inert atmosphere for 16hours. The usual work up followed by partial purification bychromatography gave impure compound 557-MS-4 (1 g, approximately 22%).

[0515] Step 2

[0516] A solution of impure compound 557-MS-4 (380 mg, assumed tocontain 0.89 mmol) in dichloromethane (5 mL) was cooled to −78° C. andtreated with a 1M solution of diisobutylaluminum hydride indichloromethane (1.79 mL, 1.79 mmol). The reaction mixture was allowedto warm to room temperature and worked up in the usual manner. Partialpurification by chromatography gave impure compound 557-MS-10 (which wasused in the next stage without further purification).

[0517] Step 3

[0518] To a solution of triphenylphosphine (910 mg, 3.47 mmol) in drydichloromethane (6 mL), at 0° C. under an inert atmosphere, was addeddrop wise a solution of carbon tetrabromide (575 mg, 1.73 mmol) in drydichloromethane (3 mL). After 10 minutes a mixed solution of impurecompound 557-MS-10 (assumed to contain 0.86 mmol) plus triethylamine(0.181 mL, 1.3 mmol) in dry dichloromethane (3 ml) was added drop wise.The reaction mixture was worked up in the usual manner to give compound557-MS-14 (331 mg, 72% from 557-MS-4).

[0519] Step 4

[0520] To a solution of compound 557-MS-14 (331 mg, 0.61 mmol) in dryTHF (2.5 mL), at −78° C. under an inert atmosphere, was added drop wisea 1.6M solution of n-butyllithium in hexanes (0.771 mL, 1.23 mmol). Thereaction mixture was warmed momentarily to 0° C. then re-cooled to −78°C. A solution of compound 480-XYL-075 (247 mg, 0.51 mmol) in drytetrahydrofuran (2.5 mL) was added drop wise. Warming to 0° C. followedby the usual work up and chromatographic purification gave compound557-MS-19 (300 mg, 57%).

[0521] Step 5

[0522] A solution of compound 557-MS-19 (300 mg, 0.35 mmol) in hexane(10 mL) was hydrogenated, at room temperature and 1 atmosphere pressure,in the presence of Lindlar's catalyst (60 mg) and quinoline (4 μL) for40 hours (fresh Lindlar catalyst (120 mg) added after 2 hours).Filtration and concentration in vacuo gave compound 557-MS-22 (302 mg)which was used in the next stage without purification.

[0523] Step 6

[0524] A solution of crude compound 557-MS-22 (assumed to contain 0.35mmol) in 1,2-dichloroethane (10 mL) was treated with benzoyl chloride(0.122 mL, 1.05 mmol), triethylamine (0.293 mL, 2.1 mmol) andN,N-4-dimethylaminopyridine (21 mg, 0.175 mmol). The usual work upfollowed by chromatographic purification gave compound 557-MS-26 (177mg, 53% from compound 557-MS-19).

[0525] Step 7

[0526] A solution of compound 557-MS-26 (177 mg, 0.183 mmol) in THF (3mL) was treated with a 1M solution TBAF in THF (1.83 mL). The usual workup gave crude compound 557-MS-43, which was used in the next stagewithout further purification (m/z: 623.3 [M-1, 24%], 249.1 [100%]).

[0527] Step 8

[0528] A solution of crude compound 557-MS-43 (84 mg, 0.13 mmol) in1,2-dichloroethane (90 mL) was added slowly to a heated solution (80°C.) of 2-chloro-1-methylpyridinium iodide (210 mg, 0.81 mmol) andtri-n-butylamine (0.192 ml, 0.81 mmol) in dichloromethane (90 mL). Theusual work up and chromatographic purification gave compound 557-MS-70(15.7 mg, 20% from compound 557-MS-26).

[0529] Step 9

[0530] A solution of compound 557-MS-70 (15.7 mg, 25 μmol) in a mixtureof ethanol (1.8 mL) and THF (0.9 mL) was treated with 1M aqueous NaOH(0.3 mL) and stirred for approximately 16 hours at room temperature. Theusual work up gave compound 557-MS-74 (6 mg, 46%).

[0531] Step 10

[0532] A solution of compound 557-MS-74 (9 mg, 18 μL) in dichloromethane(3 mL) was treated with PCC (58 mg, 0.269 mmol) in the presence ofpowdered 4 Å molecular sieves (58 mg). The reaction mixture was stirredvigorously for 90 minutes at room temperature. Basification with excesstriethylamine, followed by partial chromatographic purification gaveimpure compound 557-MS-77, which was used in the next stage withoutfurther purification (m/z: 523.1 [M+23, 100%], 365.1 [22%]).

[0533] Step 11

[0534] A solution of impure compound 557-MS-77 (1 mg, assumed to contain2 μmol) in a mixture of acetonitrile (200 μl) and dichloromethane (50μl) was treated with 48% aqueous hydrofluoric acid (50 μl). After 25minutes the usual work up followed by chromatographic purification gavecompound ER-804003 (0.3 mg, approximately 4% from compound 557-MS-74).

[0535] Preparation of Compound ER-803924 (C4 Benzyl)

[0536] Step 1

[0537] To a solution of freshly prepared LDA (0.053 mmol) in dry THF (30mL), at −78° C. under an inert atmosphere, was added drop wise asolution of methyl (S)-3-hydroxybutanoate (3 g, 0.025 mol) in dry THF(30 mL). After 2 hours at −78° C. the reaction mixture was treated dropwise with benzyl bromide (9.06 mL, 0.076 mol), then warmed to roomtemperature. The usual work up followed by chromatographic purificationgave compound 501-MS-226 (1.28 g, 24%).

[0538] Step 2

[0539] A solution of compound 501-MS-226 (1.28 g, 6.14 mmol) in DMF (10mL) was treated with tert-butyldiphenylsilyl chloride (1.76 mL. 6.75mmol) and imidazole (461 mg, 6.75 mmol) then heated at 50° C. for 4hours. The usual work up followed by chromatographic purification gavecompound 501-MS-251 (1.87 g, 68%).

[0540] Step 3

[0541] To a solution of compound 501-MS-251 (1.37 g, 3.06 mmol) in drydichloromethane (50 mL), at −78° C. under an inert atmosphere, was addeda 1.5M solution of DIBAL-H in toluene (5.11 mL, 7.66 mmol). The reactionmixture was then warmed to 0° C. and stirred at 0° C. for 2 hours. Theusual work up followed by chromatographic purification gave compound501-MS-255 (1.06 g, 83%).

[0542] Step 4

[0543] To a solution of oxalyl chloride (0.314 mL, 3.6 mmol) in drydichloromethane (8 mL), at −78° C. under an inert atmosphere, was addeddrop wise a solution of dimethylsulfoxide (0.51 mL, 7.2 mmol) in drydichloromethane (4 mL). After 30 minutes a solution of compound501-MS-255 (1.369 g, 3.27 mmol) in dry dichloromethane (8 mL) was addeddrop wise. The reaction mixture was stirred at −78° C. for 1 hour thentreated with triethylamine (2.28 mL, 16.35 mmol) then warmed slowly toroom temperature. The usual work up gave compound 501-MS-257 (1.24 g,91%).

[0544] Step 5

[0545] To a solution of triphenylphosphine (2.57 g, 9.81 mmol) in drydichloromethane (16 mL), at 0° C. under an inert atmosphere, was addeddrop wise a solution of carbon tetrabromide (1.63 g, 4.91 mmol) in drydichloromethane (8 mL). A mixed solution of compound 501-MS-257 (1.24 g,2.97 mmol) and triethylamine (0.501 ml, 3.6 mmol) was added drop wiseand the reaction mixture warmed to room temperature. The usual work upfollowed by chromatographic purification gave compound 501-MS-261 (1.47g, 79% from 501-MS-255).

[0546] Step 6

[0547] To a solution of compound 501-MS-261 (1.47 g, 2.57 mmol) in dryTUF (10 mL), at −78° C. under an inert atmosphere, was added drop wise a1.6M solution of n-butyllithium in hexanes (3.21 mL, 5.14 mmol). Thereaction mixture was warmed momentarily to 15° C. then re-cooled to −78°C. A solution of compound 480-XYL-075 (950 mg, 1.976 mmol) in dry THF(10 mL) was added drop wise. Warming to 0° C. followed by the usual workup and chromatographic purification gave compound 501-MS-265 (1.11 g,63%).

[0548] Step 7

[0549] A solution of compound 501-MS-265 (1.11 g, 1.24 mmol) in hexane(70 mL) was hydrogenated, at room temperature and 1 atmosphere pressure,in the presence of Lindlar's catalyst (350 mg) and quinoline (70 μL),for 5 hours. Filtration and concentration in vacuo gave compound501-MS-267 (1.13 g, assumed quantitative), which was used in the nextstage without purification.

[0550] Step 8

[0551] A solution of compound 501-MS-267 (160 mg, 0.178 mmol) in THF (1mL) was treated with a 1M solution of TBAF in THF (0.89 mL). Afterapproximately 15 hours the usual work up gave crude compound 501-MS-279(81 mg, approximately 82%).

[0552] Step 9

[0553] A solution of compound 501-MS-279 (81 mg, 0.146 mmol) in1,2-dichloroethane (100 mL) was added slowly to a heated solution (85°C.) of 2-chloro-1-methylpyridinium iodide (223 mg, 0.87 mmol) andtri-n-butylamine (0.207 ml, 0.87 mmol) in dichloromethane (100 mL). Theusual work up and chromatographic purification gave compound 501-MS-282(15 mg, 19%) (m/z: 555.4 [M-1; 100%], 511.4 [28%]).

[0554] Step 10

[0555] A solution of compound 501-MS-282 (15 mg, 0.027 mmol) indichloromethane (3 mL) was treated with PCC (72 mg, 0.334 mmol) in thepresence of powdered 4 Å molecular sieves (72 mg). The reaction mixturewas stirred vigorously for 90 minutes at room temperature. Basificationwith excess triethylamine, followed by chromatographic purification gavecompound 501-MS-284 (12 mg, 80%).

[0556] Step 11

[0557] A solution of compound 501-MS-284 (9 mg, approximately 0.016mmol) in a mixture of acetonitrile (1.2 mL) and dichloromethane (300 μL)was treated with 48% aqueous hydrofluoric acid (300 μL). After 20minutes the usual work up followed by chromatographic purification gavecompound ER-803924 (7.5 mg, quantitative).

[0558] Preparation of C3-Hydrogen with C4-Methyl Analogs:

[0559] Synthesis of ER-804035:

[0560] To the methyl (S)-(+)-3-hydroxy-2-methylpropionate (14.4 g, 0.121mol) in DMF (26 mL) at 0° C. were added 4-dimethylaminopyridine (0.79 g,0.006 mol), imidazole (14.3 g, 0.210 mol) andt-butyldimethylchlorosilane (23.8 g, 0.158 mol). The mixture was stirredat 0° C. for 10 min then at room temperature overnight. The mixture waspartitioned between ether and saturated sodium bicarbonate solution. Thetwo layers were separated and the aqueous layer was extracted with etherthree times. The ether extracts were combined, dried with sodium sulfateand concentrated. The crude sample was chromatographed on a silica gelcolumn eluting with 5% ethyl acetate in hexanes to get 29.6 g (98%) ofproduct, 480-XYL-073 with satisfactory ¹H-NMR data.

[0561] The compound 480-XYL-073 (5.04 g, 21.7 mmol) was dissolved inmethylene chloride (216 mL) and cooled to −78° C. To this was addeddiisobutylaluminium hydride solution in methylene chloride (1.0M, 22 mL)at a rate of 13.3 mL per hour via syringe pump down the inner side offlask wall. After completion of addition, the mixture was stirredadditional 30 min. The reaction was quenched slowly with methanol downthe wall and added some saturated aqueous solution of potasium sodiumtartrate. The mixture was warmed up to rt., added additional potasiumsodium tartrate solution and stirred vigorously for 1 h. The layers wereseparated and aqueous layer was extracted two times with methylenechloride. The combined organic layers were dried with sodium sulfate,filtered and concentrated to get 4.36 g of crude material, 480-XYL-077.The ¹H-NMR data analysis showed the desired product aldehyde: startingmaterial:over-rudeced alcohol ratio was 3:1.33:1. This mixture wasdirectly used for the synthesis of 480-XYL-079.

[0562] The compound 480-XYL-079 was synthesized following the sameprocedure as the synthesis of 554-RB-228.

[0563] DMSO (0.5 mL) in methylene chloride (24 mL) was cooled to −78° C.To this, was added oxalyl chloride solution in methylene chloride (2.0M, 1.7 mL) and the mixture was stirred for 10 min at −78° C. The compound531-YW-005 (1.4 g, 2.9 mmol) in a solution of methylene chloride (3 mLand rinsed, 2×2 mL) was added via a cannula, and the mixture was stirredfor 10 min at −78° C. To this, was added triethylamine (2.5 mL) dropwise and the mixture was stirred for 1 hour at −78° C., was warmed to 0°C. The reaction was diluted with large amount of ether and washed oncewith saturated ammonium chloride solution and water (1:1), and withwater (3×). The ether layer was concentrated in vacuo and re-dissolvedin large amount of ether. This was washed two times with water and oncewith brine. The ether layer was concentrated, azeotroped with ethylacetate and toluene and dried under high vacuum to get 1.36 g (98%),480-XYL-075 which showed good purity by ¹H-NMR spectrum. This materialwas immediately used for the synthesis of 480-XYL-081.

[0564] The synthesis of 480-XYL-084 followed the same procedure as thesynthesis of 554-RB-240.

[0565] The compound 480-XYL-084 (855 mg, 1.21 mmol) was dissolved in amixture of methanol and water (5:1, 60 mL). A slurry solution ofRieke-zinc in THF (8 mL) was added and the mixture was heated to refluxwith stirring for three hours. The mixture was filtered through a plugof celite and silica gel, rising with ethyl acetate. The filtrate wasconcentrated, re-dissolved in methylene chloride and washed withsaturated ammonium chloride solution and then with saturated sodiumbicarbonate solution. The aqueous phase was back extracted two timeswith methylene chloride and once with ethyl acetate. The combinedorganic layers were dried with sodium sulfate and concentrated to get944 mg of crude material, 480-XYL-075, which showed satisfactory purityby ¹H-NMR spectrum and directly used for the synthesis of 480-XYL-092.

[0566] The synthesis of 480-XYL-092 was the same as the procedure of thesynthesis of 554-RB-242.

[0567] ER-804035 The synthesis of ER-804035 from 480-XYL-092 wasfollowed the same procedure as for the synthesis of ER-803 064.

[0568] Preparation of ER804022:

[0569] Oxalyl chloride (6.5 mL, 74.1 mmol) was dissolved in 150 mldichloromethane at −78° C. Methyl sulfoxide (10.5 mL, 148.2 mmol) wasadded. After 20 min, the solution of starting material (5.2 g, 24.7mmol) in 50 mL of dichloromethane was added at −78° C. After stirringfor 1 h at −78° C., triethylamine (31.0 mL, 222 mmol) was added and thereaction mixture was warmed up to room temperature. It was quenched withsat. ammonium chloride and extracted with ethyl acetate. Afterpurification on silica gel column, 509-HD-183 was obtained in 79% yield.

[0570] Triphenylphosphine (13.4 g, 51.2 mmol) was dissolved in 100 mLdichloromethane at 0° C. Carbon tetrabromide (8.5 g, 25.6 mmol) wasadded. After 15 min, the solution of 509-HD-183 (4.1 g, 19.7 mmol) andtriethylamine (2.8 mL, 19.7 mmol) in 50 ml of dichloromethane was added.After stirring for 30 min, the reaction mixture was triturated withpentane. After purification on silica gel column, 509-HD-184 wasobtained in 88% yield.

[0571] 509-HD-184 (553 mg, 1.52 mmol) was dissolved in 10 mL of THF at−78° C. The

[0572] solution of n-butyl lithium (2.5M , 1.33 mL) in hexane was added.After 15 min at −78° C., the solution of 531-HYW-5 in 5 ml of THF wasadded. After stirring for 30 min at −78° C., the reaction mixture waswarmed up to room temperature. It was quenched with water and extractedwith ethyl acetate. After purification on silica gel column, 509-HD-185was obtained in 95% yield.

[0573] 509-HD-185 (750 mg, 1.09 mmol) was dissolved in 40 mL of hexane.Quinoline (50 μL) and Lindlar catalyst (120 mg) were added. The reactionmixture was stirred at room temperature under H₂ balloon atmosphere for5 h. Then the catalyst was filtered away. Quantitative amount of509-HD-186 was obtained.

[0574] 509-HD-186 (861 mg, 1.09 mmol) was dissolved in 15 mL ofdichloromethane at room temperature. Triethylamine (380 μL, 2.73 mmol),benzoyl chloride (253 μL, 2.18 mmol) and catalytic amount of DAMP wereadded, respectively. After stirring for 20 h, 0.1N sodium hydroxidesolution was added and the reaction mixture was extracted with ethylacetate. The crude product was purified on silica gel column, giving509-HD-187 in 95% yield.

[0575] 509-HD-187 (813 mg, 1.03 mmol) was dissolved in a mixture of 10mL of dichloromethane and 5 mL of water. DDQ (234 mg, 1.03 mmol) wasadded. After stirring at room temperature for 1 h, the reaction mixturewas quenched with sat. sodium bicarbonate solution and extracted withethyl acetate. After purification on silica gel column, 509-HD-188 wasobtained in 48% yield.

[0576] 509-HD-188 (313 mg, 0.47 mmol) was dissolved in 15 mL ofdichloromethane at 0° C. Triethylamine (130 μL, 0.94 mmol) andmethanesulfonyl chloride (54 μL, 0.71 mmol) were added. After stirringfor 20 min, the reaction mixture was quenched with sat. sodiumbicarbonate and extracted with dichloromethane. After purification onsilica gel column, 509-HD-189 was obtained in 93% yield.

[0577] 509-HD-189 (327 mg, 0.44 mmol) was dissolved in 10 mL of DMF.Sodium azide (85 mg, 1.32 mmol) and catalytic amount oftetrabutylammonium iodide were added. After stirring at 85° C. for 2 h,the reaction mixture was diluted with ethyl acetate and washed withwater. After purification on silica gel column, 509-HD-190 was obtainedin 93% yield.

[0578] 509-HD-190 (297 mg, 0.43 mmol) was dissolved in 10 mL of THF. Thesolution of TBAF (1N, 1.3 mL) was added. The reaction mixture wasstirred at room temperature for 1 h. It was diluted with Et₂O and washedwith H₂O. After purification on silica gel column, 509-HD-191 (215 mg)was obtained in quantitative yield.

[0579] Trimethylphosphine (1N, 1.5 mL) was dissolved in a mixture of 15mL of THF and 5 mL of water at room temperature. 509-HD-191 (215 mg,0.31 mmol) was added. After stirring for 12 h, it was concentrated andazeotroped with toluene. The residue was re-dissolved in 50 mldichloromethane. EDC (593 mg, 3.1 mmol) was added. After stirring for 2h, it was diluted with water and extracted with dichloromethane. Afterpurification on HPTLC, 509-HD-197 was obtained in 30% yield.

[0580] 509-HD-197 (51 mg, 0.092 mmol) was dissolved in 5 mL of ethanol.Sodium hydroxide (1N, 0.92 mL) solution was added. The reaction mixturewas stirred for 48 h at room temperature. It was diluted with H₂O,extracted with EtOAc. After purification on HPTLC, 11.5 mg of the majordesired single isomer 509-HD-198 was obtained as colorless oil.

[0581] 509-HD-198 (10.0 mg, 0.022 mmol) was dissolved in 3 mL ofdichloromethane. Molecular sieve (4A, 48 mg) and PCC (48 mg, 0.22 mmol)were added. The reaction mixture was stirred for 48 h at roomtemperature. After purification on prep TLC, 509-HD-200 was obtained in35% yield.

[0582] 509-HD-200 (13 mg, 0.0079 mmol) was dissolved in 0.25 mL ofdichloromethane. Then hydrofluoric acid (6N, 1 mL) was added. Thereaction mixture was stirred at room temperature for 1 h. It was dilutedwith more dichloromethane, washed with water and sat. sodium bicarbonatesolution. After purification on a plug of silica gel, ER804022 wasobtained as a white solid in quantitative yield.

[0583] Preparation of ER-803027-00-01

[0584] To a magnetically stirred solution of 531-YW-2-2 (3.5 g, 13.4mmol) in 15 mL of dry DMF cooled to −0° C. (ice/water; externalthermometer) was introduced NaH (0.39 g, 16 mmol) followed by benzylbromide (0.34 g, 20 mmol). After 18 hours of stirring at roomtemperature the reaction mixture was cooled down at 0° C. and water wasadded. The reaction mixture was diluted with water and extracted withEthyl ether. The crude was purified on silica gel (Hexane/EtOAc: 90/10)to afford 447-JCH-245B (0.93 g, 20% yield).

[0585] To a magnetically stirred solution of 447-JCH-245B (0.93 g, 2.7mmol) in 8 mL of ethanol 1 M NaOH aqueous solution (4 mL) was added.After 18 hours of stirring at room temperature the reaction mixture wasdiluted with water and extracted with ethyl ether. The crude waspurified on silica gel (Hexane/EtOAc: 60/40) to afford 447-JCH-268B(0.47 g, 67% yield).

[0586] Using a procedure analogous to that described for the synthesisof intermediate 531-YW-3, 447-JCH-268B (0.450 g, 1.69 mmol) was reactedwith triphenylphosphine (0.887 g, 3.38 mmol), DEAD (0.32 mL, 2 mmol) andmethyl iodide (0.158 mL, 2.54 mmol) in toluene (16.9 mL) to afford447-JCH-271B (0.553 g, 86% yield).

[0587] To a magnetically stirred solution of 509-HD-213 (0.553 g, 1.47mmol) and 447-JCH-271B (1.06 g, 2.2 mmol) in 6 ml of a 10:1 ratio ofHMPA/THF mixture at −78° C. (dry ice/acetone; internal thermometer) wasslowly (syringe pump) introduced a 1 M solution of LiHMDS in THF (2.2mL, 2.2 mmol) diluted with 2.2 mL of the same mixture. After 30 min. at−78° C., the reaction mixture was warmed-up to 0° C. The reaction wasthen quenched by addition of a saturated aqueous solution of ammoniumChloride. The reaction mixture was diluted with water and extracted withEthyl ether. The crude was purified on silica gel (Hexane/EtOAc: 90/10to afford 447-JCH-273B (0.910 g, 84% yield).

[0588] To a magnetically stirred solution of 447-JCH-273B (0.910 g, 1.25mmol) in 24 mL of dichloromethane at 0° C. (ice/water; externalthermometer), MCPBA (0.66 g, 3.8 mmol) was added. After 15 min. ofstirring at 0° C., triethylamine was added (1.5 mL) and the reactionmixture was warmed-up to room temperature. After 45 min of stirring atroom temperature a 10% solution of sodium thiosulfate in a saturatedaqueous solution of sodium bicarbonate was added and the mixture wasstirred for 30 minutes. The reaction mixture was diluted with water andextracted with Ethyl ether. The crude was purified on silica gel(Hexane/EtOAc: 80/20 to afford 447-JCH-275B (0.70 g, 98% yield).

[0589] To a magnetically stirred solution of 447-JCH-275B (0.71 g, 1.2mmol) in 15 mL of methanol at room temperature was introduced acatalytic amount of Pd 10% on carbon. The reaction mixture was stirred18 hours at room temperature and then filtered through a pad of celite.The crude was purified on silica gel (Hexane/EtOAc: 70/30) to afford447-JCH-277B (0.58 g, 99% yield).

[0590] Using a procedure analogous to that described for the synthesisof intermediate 480-XYL-075, 447-JCH-277B (0.6 g, 1.2 mmol) was reactedwith Dess-Martin reagent (0.763 g, 1.8 mmol), and sodium bicarbonate(0.38 g) in dichloromethane (48 ml) to afford 447-JCH-280A (0.602 g).

[0591] Using a procedure analogous to that described for the synthesisof ER-803064 (stage 509-HD-108), 447-JCH-280A (0.6 g,) was reacted withintermediate 343-YW-276 (0.32 g, 1.56 mmol) in THF (18 mL) to afford447-JCH-282B (0.6 g, 70% yield from 447-JCH-277B).

[0592] Using a procedure analogous to that described for the synthesisof ER-803064 (stage 509-HD-112), 447-JCH-282B (0.6 g,) hydrogenatedusing Lindlar catalyst to afford 447-JCH-283B (0.61 g).

[0593] Using a procedure analogous to that described for the synthesisof ER-803064 (stage 509-HD-115), 447-JCH-283A (0.72 g) was reacted withbenzoyl chloride (0.37 mL, 2.63 mmol) to afford 447-JCH-285B (0.78 g,93%).

[0594] To a magnetically stirred solution of 447-JCH-285B (0.68 g, 0.86mmol) in a 2/1 dichloromethane/water mixture (26 mL) at room temperature, DDQ (0.17 g) was added. After 40 minutes of stirring at roomtemperature, the reaction mixture was diluted with ethyl acetate washedonce with an aqueous solution of sodium hydroxide (0.1N) and twice withwater. The crude product was purified by flash chromatography elutingwith hexane/ethyl acetate: (60/40) to afford 447-JCH-287B (0.52 g, 88%).

[0595] Using a procedure analogous to that described for the synthesisof ER-803064 (stage 509-HD-116), 447-JCH-287B (0.58 g, 0.86 mmol) wasreacted with TBAF (0.68 g, 2.6 mmol) in THF (2.6 mL) to afford447-JCH-288A (0.47 g).

[0596] To a magnetically stirred solution of 447-JCH-288A (0.31 g, 0.54mmol) and triphenylphosphine (0.34 g, 2.17 mmol) in THF (43 mL) at roomtemperature, DEAD (0.57 g, 2.17 mmol) was added. After 1 hour ofstirring at room temperature, the reaction mixture was concentratedunder vacuum. The crude product was purified by flash chromatographyeluting with hexane/ethyl acetate: (70/30) to afford 447-JCH-290B (0.21g, 70%).

[0597] Using a procedure analogous to that described for the synthesisof ER-803064 (stage 509-HD-119), 447-JCH-290B (0.21 g, 0.38 mmol) wasreacted with sodium hydroxide (1M solution, 1.9 mL, 1.9 mmol) in ethanol(5.7 mL) to afford 447-JCH-294B (0.128 g, 73%).

[0598] To a magnetically stirred solution of 447-JCH-294B (0.07 g, 0.155mmol) in dichloromethane (15 mL) at room temperature sodium bicarbonate(0.08 g) and Dess-Martin reagent (165 mg, 0.388 mmol) were added. After45 minutes of stirring at room temperature, a 10% (w/w) thiosulfatesolution in saturated aqueous solution of sodium bicarbonate was added.The reaction mixture was diluted with water and extracted with ethylether. The crude product was purified by flash chromatography elutingwith n-hexane/ethyl acetate: (60/40) 447-JCH-295B (0.06 g, 88%).

[0599] Using a procedure analogous to that described for the synthesisof ER-803064 (stage 509-HD-125), 447-JCH-295B (0.017 g, 0.038 mmol) wasreacted with HF (48%) (0.85 ml) in acetonitrile (3.4 mL) to afford447-JCH-296B/ER-803027 (0.006 g, 97%).

[0600] Preparation of B2329:

[0601] A mixture of 1,3-propanediol (15 g, 197 mmol), p-anisaldehydedimethylacetal (37 mL, 217 mmol) and p-toluene sulfonic acid (35 mg) wasstirred under slight vacuum for 6 hours at 35° C. in DMF (35.5 mL). Thereaction mixture was cooled to room temperature, and then a saturatedaqueous solution of sodium bicarbonate was added. The reaction mixturewas diluted with water and extracted with ethyl acetate to afford447-SG-089A (35.8 g). The crude was used directly into the next stepwithout purification.

[0602] To a magnetically stirred solution of 413-SG-89A (12.95 g, 66.67mmol) in dichloromethane (225 mL) cooled to −5° C. (Ice/salt, internalthermometer), 1 M solution of DIBAL-H in toluene (100 mL, 100 mmol) wasadded. After 2 hours of stirring at room temperature, the reaction wasquenched by addition of methanol (100 mL). After 2 hours of vigorousstirring, 100 ml of a saturated aqueous solution of sodium sulfate wasadded. After 1 hour of stirring, the reaction mixture was diluted with100 mL of ethyl ether and stirred at room temperature for half an hour.The reaction mixture was filtered through a plug of celite and thesolvent was removed by evaporation. The crude product was purified byflash chromatography eluting with hexane/ethyl acetate (2:1 then 1/1) toafford 447-SG-89B (11.45 88% yield).

[0603] To a magnetically stirred solution of 413-SG-89B (2 g, 10.9 mmol)in dichloromethane (225 mL) cooled to 0° C. (Ice/water, externalthermometer) DMSO (2.5 mL, 35.67 mmol) was added, followed by P₂O₅ (5.06g, 35.67 mmol). After one hours of stirring at room temperature thereaction was cooled down to 0° C. and triethylamine (7.1 mL, 50.95 ml)was added. After 45 minutes of stirring at room temperature. Thereaction mixture was diluted with water and extracted withdichloromethane. The solvent was removed by evaporation. The residue wastriturated with ether and the solid was filtered-off and wash withether. The solvent was removed by evaporation to afford 413-SG-106A (2.1g), the crude was used directly into the next step without purification.

[0604] Using a procedure analogous to that described for the synthesisof intermediate 343-YW-276, 413-SG-106A (10.9 mmol) was reacted withtriphenylphosphine (7 g, 26.49 mmol), carbon tetrabromide (4.39 g, 13.25mmol) and triethylamine (1.4 mL, 10.9 mmol) in dichloromethane (12.6mL). The crude product was purified by flash chromatography eluting withpentane/dichloromethane (1/1) to afford 413-SG-106B (3.04 g, 85% yield).

[0605] Using a procedure analogous to that described for the synthesisof intermediate 554-RB-240B, 413-SG-106B (1.66 g, 4.73 mmol) was reactedwith n-BuLi (2.5M in toluene, 4.2 mL, 10.41 mmol) in THF (32.5 mL) at−78° C. The resulting alkynyl lithium was then reacted with intermediate343-YW-277 (1.64 g, 3.97 mmol) in THF (12 mL) at −78° C. The crudeproduct was purified by flash chromatography eluting with hexane/ethylacetate (3/1) to afford 413-SG-110B (2.01 g, 86% yield).

[0606] Using a procedure analogous to that described for the synthesisof intermediate 554-RB-241, 413-SG-110B (1.4 g, 2.27 mmol) was dissolvedin hexane (32 mL) and hydrogenated using Lindlar catalyst to afford413-SG-167A (1.4 g).

[0607] Using a procedure analogous to that described for the synthesisof 554-RB-242, 413-SG-167A (1.37 g, 2.27 mmol) was reacted with benzoylchloride (0.53 mL, 4.54 mmol), triethylamine (0.79 mL, 5.68 ml) and acatalytic amount of DMAP in dichloromethane (12 mL). The crude productwas purified by flash chromatography eluting with hexane/ethyl acetate(3/1) to afford 413-SG-169B (1.58 g, 99% yield).

[0608] 413-SG-169B (1.58 g, 2.22 mmol) was reacted with TBAF (0.88 g,3.34 mmol) in THF (6 mL) at room temperature. The reaction mixture wasdiluted with water and extracted with ethyl ether. The crude product waspurified by flash chromatography eluting with hexane/ethyl acetate (5/1and then 1/1) to afford 413-SG-169B (1.02 g, 98% yield).

[0609] Using a procedure analogous to that described for the synthesisof 554-RB-260, 413-SG-170B (1.02 g, 2.17 mmol) was reacted withtriphenylphosphine (0.97 g, 3.69 mmol), DEAD (0.36 mL, 2.28 mmol) andmethyl iodide (0.175 mL, 2.82 mmol) in toluene (19 mL). The crudeproduct was purified by flash chromatography eluting with hexane/ethylacetate (9/1 and then 5/1) to afford 413-SG-163B (1.12 g, 98% yield).

[0610] Using a procedure analogous to that described for the synthesisof 531-YW-4, 413-SG-173B (1.12 g, 1.93 mmol) was reacted withintermediate 509-HD-213 (1.2 g, 2.51 mmol) and LiHMDS (1M solution inTHF, 2.3 mL, 2.3 mmol) in a 10 to 1 THF/HMPA mixture (17.3 mL) to afford413-SG-174A. 413-SG-174A (crude) was reacted with MCPBA (0.61 g, 1.93mmol) and triethylamine (1.6 mL, 11.6 mmol). The crude product waspurified by flash chromatography eluting with hexane/ethyl acetate (5/1and then 3/1) to afford 413-SG-174B (0.895 g, 45% yield).

[0611] Using a procedure analogous to that described for the synthesisof 453-MS-262, 413-SG-174B (0.89 g, 1.14 mmol) was reacted with DDQ(0.31 g, 1.37 mmol) in a 2/1-dichloromethane/water mixture (48 mL). Thecrude product was purified by flash chromatography eluting withhexane/ethyl acetate (5/1 and then 3/1) to afford 413-SG-177B (0.454 g,61% yield).

[0612] Using a procedure analogous to that described for the synthesisof ER-803064 (stage 509-HD-116), 413-SG-177B (0.45 g, 0.691 mmol) wasreacted with TBAF (0.542 g, 2.07 mmol) in THF (2.2 ml). The crudeproduct was purified by flash chromatography withdichloromethane/methanol: (95/5) to afford 413-SG-179B (0.31 g, 79%yield).

[0613] 413-SG-179B (0.31 g, 0.54 mmol) was reacted withtriphenylphosphine (0.175 g, 0.658 mmol) and DEAD (0.105 ml, 0.658 mmol)in THF (43 ml). The crude product was purified by flash chromatographywith hexane/ethyl acetate: (3/1) to afford 413-SG-180B (0.2 g, 69%yield).

[0614] Using a procedure analogous to that described for the synthesisof ER-803064 (stage 509-HD-119), 413-SG-180B (0.18 g, 0.334 mmol) wasreacted with sodium hydroxide (1M solution, 1.7 mL, 1.7 mmol) in a 2/1mixture ethanol/THF (10 mL) to afford 413-SG-182A (0.16 g). The crudeproduct was used for the next step without further purification.

[0615] Using a procedure analogous to that described for the synthesisof ER-803064 (stage 509-HD-125), 413-SG-182A (0.14 g, 0.32 mmol) wasreacted with PCC (0.84 g, 3.87 mmol) in dichloromethane (34 mL) withmolecular sieves 4 Å (800 mg). The crude product was purified by flashchromatography with hexane/ethyl acetate: (70/30) to afford 413-SG-188B(0.07 g, 52% yield).

[0616] Using a procedure analogous to that described for the synthesisof ER-803064 (final step), 413-SG-188B (0.067 g, 0.157 mmol) was reactedwith HF (6 M solution in acetonitrile, 17.7 ml) in dichloromethane (3.1ml). The crude product was purified by flash chromatography withhexane/ethyl acetate: (60/40) to afford 413-SG-193B/B-2329 (0.05 g, 91%yield).

[0617] Preparation of B2395:

[0618] To a magnetically stirred solution of 413-SG-178B (0.194 g, 0.289mmol) and triethylamine (0.08 mL, 0.578 mmol) in dry dichloromethanecooled to 0° C. (ice/water; external thermometer) was introducedmethanesulfonyl chloride (0.034 ml, 0.434 mmol). After 1 hour ofstirring at 0° C. a saturated aqueous solution of sodium bicarbonate wasadded. The reaction mixture was diluted with water and extracted withdichloromethane. The crude was purified on silica gel (Hexane/EtOAc:1/1) to afford 413-SG-184B (0.215 g, 99% yield).

[0619] A solution of 413-SG-184B (0.216 g, 0.288 mmol), sodium azide(0.028 g, 0.432 mmol) and catalytic amount of tetra-butyl ammoniumiodide in DMF was magnetically stirred at 85° C. After 90 minutes thereaction mixture was concentrated under vacuum. The crude was purifiedon silica gel (Hexane/EtOAc: 5/1) to afford 413-SG-185B (0.086 g, 93%yield)

[0620] Using a procedure analogous to that described for the synthesisof ER-803064 (stage 509-HD-116), 413-SG-185B (0.19 g, 0.27 mmol) wasreacted with TBAF (0.21 g, 0.8 mmol) in THF (1 ml) to afford 413-SG-186A(0.14 g). The crude product was used in the next step withoutpurification.

[0621] To a magnetically stirred solution of 413-SG-186A (0.092 g, 0.156mmol) in THF/water 4/1 (1.5 mL) at room temperature was introducedtrimethylphosphine (0.78 mL, 0.778 mmol). After 18 hours of stirring atroom temperature the reaction mixture was concentrated under vacuum. Theresidue was diluted with water and extracted with dichloromethane toafford 413-SG-217A. The crude was dried and used in the next stepwithout purification.

[0622] To a magnetically stirred solution of 413-SG-217A (0.156 mmol) indichloromethane (0.2 mL) at room temperature was introduced EDC (0.10mg, 0.504 mmol). After 4 hours of stirring at room temperature thereaction mixture was Concentrated under vacuum. The crude was purifiedon silica get (Hexane/EtOAc: 1/1) to afford 413-SG-217B (0.036 g, 42%yield).

[0623] Using a procedure analogous to that described for the synthesisof ER-803064 (stage 509-HD-119), 413-SG-217B (0.036 g, 0.065 mmol) wasreacted with sodium hydroxide (1M solution, 0.3 mL, 0.3 mmol) in a 2/1mixture of ethanol/THF (2 mL) to afford 413-SG-221A (0.031 g). The crudeproduct was used for the next step without further purification.

[0624] Using a procedure analogous to that described for the synthesisof ER-803027 (stage 447-JCH-295), 413-SG-221A (0.014 g, 0.031 mmol) wasreacted with Dess-Martin reagent (80 mg, 0.188 mmol) and 2.6-lutidine(0.036 mL, 0.313 mmol) in dichloromethane (2.1 mL). The crude waspurified on silica gel (dichloromethane/methanol: 98/2) to afford413-SG-226A (0.005 g, 36% yield).

[0625] Using a procedure analogous to that described for the synthesisof ER-803064 (final step), 413-SG-226AB (0.01 g, 0.022 mmol) was reactedwith HF (1.5 M solution in acetonitrile, 5 mL) in dichloromethane (2mL). The crude product was purified by flash chromatography eluting withn-hexane/ethyl acetate: (3/1) to afford 413-SG-235B (0.004 g, 50%yield).

[0626] Preparation of C8-Deoxy Analogs, NF0530, NF0531, NF0552 andNF0761

[0627] L-Dimethylmalate (50 g, 308.4 mmol) was dissolved in dry THF (308mL) and cooled to 0° C. Then BH₃-Me₂S complex (10M, 1.1 eq., 34 mL, 0.34mol) was added dropwise and then the mixture was allowed to warm to rt.After stirred for 90 min then re-cooled to 0° C., NaBH₄ (0.05 eq., 15.4mmol, 583 mg) was added and stirred for additional 60 min. The reactionwas quenched with MeOH, and concentrated under reduced pressure. Thecrude residue was purified by chromatography on silica gel (AcOEt/MeOH)to afford MK-001 (26 g, 63%).

[0628] To a solution of MK-001 (10.0 g, 74.6 mmol) and p-anisaldehydedimethyl acetal (16.5 mL, 96.9 mmol) in 150 mL of dry CH₂Cl₂ was addedDL-10-CSA (35 mg, 0.15 mmol) at 0° C., then the reaction mixture wasallowed to warm to rt gradually. After 1 day 0.042 mL of Et₃N was addedthen evaporated. The crude product was purified by chromatography onsilica gel (Hexane/AcOEt: 5/1 to 3/1) to afford MK-002 (12.1 g, 64%).

[0629] MK-002 (12.1 g, 48.0 mmol) was dissolved in dry CH₂Cl₂-DME (240mL-240 mL) and cooled to −78° C. Then DIBAL-H in hexane (1.0 M , 50.4mL, 50.4 mmol) was added dropwise over 30 min and the mixture wasstirred for additional 100 min at −78° C. The reaction was quenched withMeOH (6 mL) then poured into a stirred solution of AcOEt and aqueoussaturated Na/K tartrate. The organic extract was washed with brine,dried over Na₂SO₄, filtered and concentrated to afford crude oil ofMK-003 (11.84 g), which was used for next step without purification.

[0630] Ph₃PCH₃ ⁺Br⁻ (34.3 g, 96.0 mmol) was dissolved in dry THF (320mL). The mixture was cooled to 0° C., and n-BuLi in hexane (1.6 M , 51.0mL, 81.6 mmol) was added slowly. After stirring for 120 min, a solutionof the crude MK-003 (11.84 g) in 50 mL of dry THF was added slowly. Thereaction was stirred for 30 min at 0° C. and followed by overnight atrt, then quenched with saturated aqueous solution of NH₄Cl. The mixturewas extracted with AcOEt, washed with brine, dried over MgSO₄, filteredand concentrated to give crude oil. The crude oil was diluted withEt₂O-hexane, the generated precipitate was filtered off, then thefiltrate was evaporated. The residual oil was purified by chromatographyon silica gel (Hexane/AcOEt: 20/1 to 8/1) to afford oil of MK-004 (4.25g, 40% 2 steps).

[0631] To a solution of MK-004 (4.05 g, 18.4 mmol) in 92 mL of dry THFwas slowly

[0632] added BH₃-Me₂S (2.0 M in THF, 4.60 mL, 9.2 mmol) at 0° C., thenthe reaction mixture was stirred at 0° C. for 90 min after which it wasallowed to warm to rt and stirred for 120 min. The solution wasre-cooled to 0° C., then treated with aqueous 3N-NaOH (28 mL) andaqueous 30%-H₂O₂ (28 mL) with vigorous stirring. The mixture wasextracted with Et₂O, washed with saturated aqueous solution of Na₂SO₃,dried over MgSO₄, filtered and concentrated to gave crude oil. The crudeoil was purified by chromatography on silica gel (Hexane/AcOEt: 1/1 to2/3) to afford oil of MK-005 (2.64 g, 60%).

[0633] Using similar procedure for the synthesis of the TBS-ether,intermediate of TM-03, from 491-HAD-46, MK-005 (2.64 g, 11.1 mmol) wasconverted to crude MK-006 (3.91 g). It was used for next step withoutpurification.

[0634] MK-006 (3.91 g) was dissolved in 74 mL of dry CH₂Cl₂ and cooledto −78° C. DIBAL-H in hexane (1.0 M , 5 eq., 55.3 mL, 55.3 mmol) wasadded dropwise over 30 min, and the solution was stirred at −78° C. foradditional 60 min after which it was allowed to warm to 0° C. over 50min. The reaction was quenched with MeOH (7 mL) then poured into astirred solution of AcOEt and saturated aqueous Na/K tartrate. Theorganic extract was washed with brine, dried over Na₂SO₄, filtered andconcentrated to give a crude oil. The crude oil was purified bychromatography on silica gel (Hexane/AcOEt: 3/1) to afford an oil ,MK-007 (3.19 g, 81% 2 steps).

[0635] To a solution of (COCl)₂ (3 eq., 2.24 mL, 25.6 mmol) in 84 mL ofdry CH₂Cl₂, DMSO (6 eq., 3.64 mL, 51.3 mmol) was added slowly at −78° C.After 15 min at −78° C., a solution of MK-007 (3.03 g, 8.55 mmol) wasadded dropwise into the reaction at −78° C. After

[0636] 30 min at that temperature, Et₃N (9 eq., 10.7 mL, 76.9 mmol) wasadded slowly. The reaction mixture was allowed to warn to −10° C.gradually. It was quenched with saturated aqueous solution of NH₄Cl,extracted with AcOEt-hexane, washed with aqueous solution of KHSO₄ thenbrine, dried over Na₂SO₄, filtered and concentrated to gave crude oil ofMK-008 (3.52 g). It was used for next step without purification.

[0637] Using similar procedure for the synthesis of TM-04 from TM-03 andTM-02, MK-008 (3.52 g) was coupled with NY-22 (6.06 g) and thenconverted to pure MK-009 (5.76 g, 100% 2 steps) as mixture ofdiastereomers on propargylic position.

[0638] Using similar procedure for the synthesis of TM-05 from TM-04,MK-009 (5.75 g) was converted to crude MK-010 (6.11 g) as mixture ofdiastereomers on allylic position.

[0639] Using similar procedure for the synthesis of 554-RB-242 from554-RB-241, crude MK-010 (6.11 g) was converted to pure MK-011 (5.93 g,89% 2 steps) as mixture of diastereomers on allylic position.

[0640] To a stirred solution of MK-011 (5.93, 7.59 mmol) in 76 mL of99.5% EtOH, PPTS (0.15 eq., 286 mg, 1.14 mmol) was added at rt, then themixture was warmed to 45° C. After 1 day it was diluted with AcOEt, thenwashed with saturated aqueous solution of NaHCO₃ and brine, dried overMgSO₄, filtered and concentrated to give crude oil. The crude oil waspurified by chromatography on silica gel (Hexane/AcOEt: 2/1) to affordoil of MK-012 (4.94 g, 98%).

[0641] Using similar procedure for the synthesis of 554-RB-260 from554-RB-244, MK-012 (4.20 g, 6.30 mmol) reacted with DIAD and methyliodide in the presence of Ph₃P to give pure MK-013 (4.74 g, 97%) asmixture of diastereomers on allylic position.

[0642] To a stirred mixture of diphenol (13.0 g, 66.3 mmol), MeOH (6.2mL, 152 mmol), and iPr₂EtN (13.9 mL, 79.5 mmol) in 110 mL of CH₃CN,TMSCHN₂ in hexane (2M , 38.1 mL, 76.2 mmol) was added dropwise over 80min at rt, then stirred overnight. The reaction was quenched with 5%citric acid aqueous solution and extracted with AcOEt. The organicextract was washed with saturated aqueous solution of NaHCO₃ and brine,dried over MgSO₄, filtered and concentrated. The crude product waspurified by chromatography on silica gel (Hexane/AcOEt: 9/1) to affordcrystals of MK-014 (12.4 g, 89%).

[0643] Using similar procedure for the synthesis of 509-HD-209 from509-HD-207, MK-014 (5.2 g, 25 mmol) was converted to pure MK-015 (6.3 g,100%).

[0644] Using similar procedure for the synthesis of 509-HD-211 from509-HD-209, MK-015 (6.0 g, 23.5 mmol) was converted to a mixture ofMK-016 (5.3 g, 55%) and inseparable diselenide (2.9 g, 22%)

[0645] Using similar procedure for the synthesis of compound 4 fromcompound 2 and compound 3, MK-013 (1.5 g, 1.93 mmol) was coupled withmixture of MK-016 and diselenide (including 2.90 mmol of MK-016) toafford crude oil of MK-017 (4.3 g). It was used for next step withoutpurification.

[0646] Using similar procedure for the synthesis of compound 5 fromcompound 4, crude MK-017 (4.3 g) was converted to MK-018 (1.60 g, 92% 3steps) as compound purified.

[0647] MK-018 (1.59 g, 1.76 mmol) was dissolved in 25 mL of THF. Then,tetrabutylammonium fluoride (TBAF) in THF (1M, 7.0 mL, 7.0 mmol) wasadded at rt. The mixture was stirred for 38 hrs before saturated aqueoussolution of NH₄Cl was added. The mixture was extracted with AcOEt andthe organic extract was washed with brine, dried over Na₂SO₄, filteredand concentrated. The crude product was purified by chromatography onsilica gel (hexane/AcOEt: 5/3 to 1/2) to afford oil of MK-019 (1.10 g,93%).

[0648] To a stirred solution of MK-019 (1.07 g, 1.61 mmol) in 20 mL ofEtOH was added 32 mL of aqueous 1N-NaOH, then the mixture was warmed to100° C. After 32 hrs, it was quenched with 32 mL of aqueous 1N-HCl andextracted with AcOEt. The organic extract was washed with brine, driedover Na₂SO₄, filtered and concentrated. The crude product was purifiedby chromatography on silica gel (AcOEt/MeOH: 9/1) to afford oil ofMK-020 (860 mg, 100%).

[0649] Using similar procedure for the synthesis of TM-12 from TM-11,MK-020 (860 mg, 1.61 mmol) was converted to mixture of MK-021 and MK-022(402 mg, 49% 2 steps; MK-021:M K-022=85:15).

[0650] To a stirred suspension of Dess-Martin periodinane (1.01 g, 2.38mmol) in 40 mL of dry CH₂Cl₂, a solution of MK-021 and MK-022 (402 mg,0.794 mmol) in 40 mL of dry CH₂Cl₂ was added at 0° C., then the mixturewas warmed to rt. After 14 hrs it was re-cooled to 0° C., diluted withAcOEt, washed with saturated aqueous solution of Na₂SO₃, NaHCO₃ andbrine, dried over Na₂SO₄, filtered and concentrated to give crude oil.The crude oil was purified by chromatography on silica gel(Hexane/AcOEt: 3/1) to afford colorless crystals of NF0552 (301 mg, 74%)and colorless oil of NF0530 (35 mg, 9%).

[0651] Using similar procedure for the synthesis of NF0675 from TM-13,NF0552 (263 mg, 0.515 mmol) was converted to NF0530 (199 mg, 83%) ascompound purified.

[0652] To a stirred mixture of NF0530 (233 mg, 0.499 mmol) in 17 mL ofCH₂Cl₂ and 1.7 mL of aqueous phosphate buffer (pH 6.86) was portionwiseadded DDQ (283 mg, 1.25 mmol) at0° C., then the mixture was allowed towarm to rt slowly. After 3.5 hrs it was quenched with aqueous solutionof NaHCO₃ and diluted with AcOEt. The organic extract was washed withaqueous solution of NaHCO₃ and brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified by chromatography on silicagel (hexane/AcOEt: 5/2) to afford colorless crystals of NF0531 (143 mg,83%).

[0653] NF0552 (30 mg, 0.059 mmol) was treated with DDQ (3 eq.) at rtusing similar procedure for the synthesis of NF0531 from NF0530. Themessy reaction was worked up in the usual manner. Purification bychromatography on silica gel (Hexane/AcOEt: 1/3) gave colorless oil ofNF0761 (1.7 mg, 7%). NF0761 was analyzed by HRMS; FAB+ m/z407(MH+),Anal. Calcd for C21H26O8: MH+, 407.1706 Found 407.1711(MH+).

[0654] Preparation of C11-C12, Cyclopropyl Analogs, NF1226 and NF1227

[0655] Using the same procedure for the synthesis of TM-03 from531-yw-2-3 (491-HAD-46), MK-023 was obtained.

[0656] To a stirred mixture of MK-023 (2.5 g, 8.61 mmol) and 4-MPMCl(1.63 mL, 12.0 mmol) in 40 mL of DMF, NaH (66%, 344 mg, 9.47 mmol) wasadded portionwise at 0° C. and the mixture was warmed to rt. Afterstirred for 3 hrs, the reaction was quenched with saturated aqueoussolution of NH₄Cl and extracted with AcOEt. The organic extract waswashed with saturated aqueous solution of NaHCO₃ and brine, dried overNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel (Hexane/AcOEt: 15/1) to afford colorlessoil of the fully protected tetraol (2.83 g, 80%).

[0657] The fully protected tetraol (3.11 g, 7.58 mmol) was dissolved in38 mL of THF. Then, tetrabutylammonium fluoride (TBAF) in THF (1M, 9.9mL, 9.9 mmol) was added at rt. The mixture was stirred for 2 hrs beforesaturated aqueous solution of NH₄Cl was added. The mixture was extractedwith AcOEt and the organic extract was washed with brine, dried overNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel (Hexane/AcOEt: 1/1) to afford oil of MK-024(2.22 g, 99%).

[0658] Using similar procedure for the synthesis of 531-YW-3 from531-YW-2-3, MK-024 (1.90 g, 6.41 mmol) was converted to MK-025 (2.36 g,90%) as compound purified.

[0659] Using similar procedure for the synthesis of 531-YW-4, MK-025(2.36 g, 5.80 mmol) coupled with 509-HD-213 (3.63 g, 7.54 mmol) wasconverted to MK-026 (3.10 g, 89% 3 steps) as compound purified.

[0660] To a stirred solution of MK-026 (2.0 g, 3.32 mmol) in 130 mL oftoluene were added Et₂Zn in hexane (1M, 16.6 mL, 16.6 mmol) and CH₂I₂(1.34 mL, 16.6 mmol) at −30° C. After stirred for 30 min at −30° C., itwas warmed to rt gradually over 2 hrs and then quenched with saturatedaqueous solution of NH₄Cl. (Note: To avoid decomposition of targetproduct, short reaction time, 1˜2 hrs, was required regardless ofconversion rate.). The mixture was extracted with AcOEt and the organicextract was washed with brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified by chromatography on silicagel (Hexane/AcOEt: 6/1 to 5/1) to afford oil of MK-027 (369 mg, <19%)including small amount of MK-026, and MK-026 (519 mg, <26%) includingsmall amount of MK-027 was recovered. The recovered MK-026 was treatedagain in the same manner to give MK-027 (367 mg, <29%) including smallamount of MK-026. Trans cyclopropane MK-027 was obtained as mixture ofinseparable stereoisomers (1:1).

[0661] MK-027 (736 mg, ca 1.19 mmol) including 10% of MK-026 wasdissolved in 24 mL of THF and 4 mL of water. OsO₄ in ^(t)BuOH (3 w/v %,0.05 eq., 0.51 mL, 0.06 mmol), N-methylmorpholine (0.2 eq., 0.026 mL,0.24 mmol) and NaClO₃ (0.4 eq., 51 mg, 0.48 mmol) were added into thestirred solution at rt. After 2 days, to the mixture were added Celite,AcOEt, and aqueous solution of Na₂SO₃. The suspension was filtered, andthe filtrate was washed with brine, dried over Na₂SO₄, filtered andconcentrated to give crude oil of MK-027 (768 mg) which included noMK-026. It was used for next step without purification.

[0662] Using similar procedure for the synthesis of NF0531 from NF0530,the crude oil of MK-027 (768 mg) was converted to MK-028 (523 mg, 88% 2steps) as compound purified.

[0663] Using similar procedure for the synthesis of MK-009 from MK-007,MK-028 (204 mg, 0.411 mmol) was converted to MK-029 (231 mg, 69% 2steps) as compound purified.

[0664] MK-029 (231 mg, 0.283 mmol) was dissolved in 7 mL of 99.5% EtOHand 7 mL of hexane (note: no reaction in hexane). Then, quinoline (0.3eq., 0.01 mL, 0.085 mmol) and 5% Pd-BaSO₄ on carbon (0.05 eq., 30 mg,0.014 mmol) were added. H₂ balloon was mounted and the mixture waspurged with H₂. After stirred for 3.5 hrs under H₂ (1 atm) at rt, thereaction mixture was filtered through Celite and the filtrate wasevaporated to give a crude oil of cis-olefin (240 mg).

[0665] Using similar procedure for the synthesis of 554-RB-242 from554-RB-241, the crude cis-olefin (240 mg) was converted to MK-030 (216mg, 83% 2 steps) as compound purified.

[0666] Using similar procedure for the synthesis of MK-019 from MK-018,MK-030 (215 mg, 0.233 mmol) was directly converted to a carboxylic acidMK-031 (125 mg, 92%) as compound purified.

[0667] Using similar procedure for the synthesis of TM-12 from TM-11,MK-031 (123 mg, 0.210 mmol) was converted to crude lactonized product.It was purified by chromatography on silica gel (Hexane/AcOEt: 4/1, 3/1,to 1/1) to afford oil of MK-032 (27 mg, 22%) and oil of des-MOM formMK-033 (20 mg, 18%).

[0668] To a stirred solution of MK-032 (24 mg, 0.0424 mmol) in 0.85 mLof EtOH and 0.85 mL of THF was added aqueous 1N NaOH (2.1 eq., 0.089 mL,0.0889 mmol). After stirred for 3 days at rt, the mixture was dilutedwith AcOEt and washed with brine, dried over Na₂SO₄, filtered andconcentrated to give crude product of MK-034 (24 mg). It was used fornext step without purification.

[0669] The crude MK-034 (24 mg, assumed to contain 0.0424 mmol) wasdissolved in 9 mL of CH₂Cl₂. To the solution were added Molecular sieve4A (45 mg) and PDC (3 eq., 48 mg, 0.127 mmol) at rt. The reactionmixture was stirred for 4 days at rt, then diluted with Et₂O and passedthrough a pad of Celite. The filtrate was evaporated to give crudeproduct. It was purified by chromatography on silica gel (Hexane/AcOEt:3/1 to 2/1) to afford colorless oil of MK-035 (less polar single isomeron trans cyclopropane, 3.6 mg, 18% 2 steps), colorless oil of MK-036(polar single isomer on trans cyclopropane, 5.8 mg, 30% 2 steps) andpale yellow oil of MK-037 (isomerized trans olefin, 5.4 mg, 28% 2steps).

[0670] 50% Hydrofluoric acid (24N, 0.2 mL) was added to less polarisomer MK-035 (3.6 mg, 0.00782 mmol) in 0.8 mL of CH₃CN and stirred for1 hr at 0° C. After stirred at rt for additional 1 hr, the reactionmixture was quenched with saturated aqueous solution of NaHCO₃ andextracted with AcOEt. The organic extract was washed with brine, driedover MgSO₄, filtered and concentrated to crude product. It was purifiedby chromatography on silica gel (hexane/AcOEt: 1/2) to afford colorlesscrystals of NF1226 (2.3 mg, 59%) as single isomer.

[0671] Using similar procedure for the synthesis of MK-034 from MK-032,MK-033 (20 mg, 0.0383 mmol) was converted to diol intermediate (13 mg,81%).

[0672] Using similar procedure for the synthesis of NF530 from MK-022,the diol intermediate (15 mg, 0.0358 mmol) was converted to MK-038 (4.7mg, 32%).

[0673] Using similar procedure for the synthesis of NF0675 from TM-13,MK-038 (4.8 mg, 0.0115 mmol) was converted to colorless crystals ofNF1227 (3.6 mg, 83%, single isomer).

[0674] Using similar procedure for the synthesis of NF1226 from MK-035,polar isomer MK-036 (5.8 mg, 0.0126 mmol) was converted to colorlesscrystals of NF1227 (2.6 mg, 55%, single isomer).

[0675] NF1227 differs from NF1226 as to stereochemistry on transcyclopropane.

[0676] Preparation of C11-C12 Amide Analogs, NF1535, NF1537, and NF2306

[0677] Exemplary Synthetic Procedure for NF1535 and NF1537

[0678] Using similar procedure for the synthesis of NY-07 from NY-06,MK-014 (12.40 g, 59.0 mmol) was converted to MK-039 (13.68 g, 92%) andpurified.

[0679] MK-039 (13.68 g, 54.2 mmol) was dissolved in 360 mL of CCl₄ andthe solution was heated up to reflux. To the stirred solution wasportionwise (over 1.5 hrs) added a mixture of NBS (11.1 g, 62.4 mmol,1.15 eq.) and (PhCO)₂O₂ (722 mg, 2.98 mmol, 0.055 eq.) and stirred foradditional 30 min at reflux. The reaction mixture was cooled to rt andinsoluble material was filtered off, then the filtrate was concentratedto give crude product. The crude product was purified by chromatographyon silica gel (hexane/AcOEt: 4/1 to 3/1) to afford colorless oil ofMK-040 (14.51 g, <81%) including small amount of starting material anddibromide. It was used for next step without further purification.

[0680] To a solution of MK-040 (14.51 g, assumed to contain 43.82 mmol)in 220 mL of DMSO was added a solution of AgBF₄ (11.09 g, 57.0 mmol) in55 mL of DMSO at rt. After 2 hrs, Et₃N (18.3 mL, 131.4 mmol) was addedand stirred for 40 min at rt. The reaction mixture was diluted withAcOEt, then washed with saturated aqueous solution of NaHCO₃ and brine,dried over Na₂SO₄, filtered and concentrated to give a crude oil. Thecrude product was purified by chromatography on silica gel(hexane/AcOEt: 3/1 to 1/1) to afford colorless oil of MK-041

[0681] To a stirred mixture of MK-041 (5.49 g, 20.63 mmol) in 140 mL of99.5% EtOH was added imidazole (421 mg, 6.19 mmol) at rt. After stiiredfor 6 days, the mixture was evaporated, diluted with AcOEt, washed withwater then brine, dried over Na₂SO₄, filtered and concentrated to givecrude colorless crystals of MK-042 (4.42 g, <96%). It was used for nextstep without purification.

[0682] To a stirred suspension of crude MK-042 (2.41 g, assumed tocontain 10.75 mmol) and K₂CO₃ (3.94 g, 28.5 mmol) in 70 mL of DMF wasadded MOMCl (1.77 mL, 23.3 mmol) at 0° C., then the mixture was allowedto warm to rt. After 14 hrs, the reaction mixture was quenched withsaturated aqueous solution of NaHCO₃ and extracted with AcOEt. Theorganic extract was washed with brine, dried over Na₂SO₄, filtered andconcentrated to give crude oil of MK-043 (2.93 g, quant.). It was usedfor next step without purification.

[0683] Crude MK-043 (2.93 g, assumed to contain 10.75 mmol) wasdissolved in 80 mL of t-BuOH and 20 mL of water. Then 2-methyl-2-butene(5.69 mL, 53.7 mmol, 5 eq.) and NaH₂PO₄-2H₂O (1.68 g, 10.75 mmol) wereadded. To the stirred suspension was portionwise added NaClO₂ (1.94 g,21.5 mmol, 2 eq.) at rt. After 1 hr at rt, the mixture was diluted withAcOEt and water, then acidified with aqueous KHSO₄ solution toapproximately pH 4. The organic extract was washed with brine, driedover Na₂SO₄, filtered and concentrated to give crude oil of MK-044 (3.20g, quant.). It was used for next step without purification.

[0684] Crude MK-044 (3.20 g, assumed to contain 10.75 mmol) wasdissolved in 72 mL of dry THF and 4.45 mL of BnOH (43.00 mmol, 4 eq.).Then Et₃N (1.80 mL, 12.90 mmol, 1.2 eq.) and DPPA (2.54 mL, 11.82 mmol,1.1 eq.) were added. The mixture was heated to 65° C. and stirred for 15hrs, then cooled to rt. The mixture was diluted with AcOEt and saturatedaqueous hsolution of NH₄Cl. The organic extract was washed withsaturated aqueous solution of NaHCO₃ then brine, dried over Na₂SO₄,filtered and concentrated to give a crude oil. The crude product waspurified by chromatography on silica gel (hexane/AcOEt: 5/1) to affordcolorless crystals of

[0685] To a stirred solution of MK-045 (2.73 g, 7.02 mmol) in 80 mL ofEtOH was added aqueous 0.5N-NaOH (1.15 eq., 16.2 mL, 8.07 mmol). Afterstirred for 2 days at rt the reaction mixture was cooled to 0° C.,quenched with aqueous 0.2N HCl (1.15 eq., 40.3 mL, 8.07 mmol) anddiluted with water (40 mL) to produce a precipitation. The precipitationwas filtered, washed with hexane-AcOEt (15 mL-2 mL) and dried underreduced pressure to afford pure colorless crystals of MK-046 (1.58 g,62%).

[0686] Ph₃P (2.98 g, 11.37 mmol, 2.6 eq.) was dissolved in 30 ml of dryTHF and cooled to 0° C. 40% DEAD in toluene (4.76 mL, 10.49 mmol, 2.4eq.) was added and stirred for 30 min at 0° C. To the stirred solutionwas dropwise added a mixture of MK-046 (1.58 g, 4.37

[0687] mmol) and 2-(trimethylsilyl)ethanol (0.94 mL, 6.56 mL, 1.5 eq.)in 25 mL of THF at 0° C. After 30 min, the reaction mixture was warmedup to rt gradually over 1 hr. The resulting mixture was evaporated andpurified by chromatography on silica gel (hexane/AcOEt: 6/1 to 5/1) toafford oil of MK-047 (2.01 g, 99%).

[0688] MK-047 (2.01 g, 4.35 mmol) was dissolved in 60 mL of AcOEt. Then10% Pd/C (50% wet, 200 mg) was added. H₂ balloon was mounted and themixture was purged with H₂ (1 atm). After stirred overnight at rt it wasworked up in usual manner and purified by chromatography on silica gel(hexane/AcOEt: 6/1 to 5/1) to afford colorless crystals of MK-048 (1.20g, 84%).

[0689] Using similar procedure for the synthesis of 343-YW-203 from(S)-1,3-butanediol, (R)-1,3-butanediol (9.80 g, 108.7 mmol) wasconverted to MK-049 (21.54 g, 94% 2 steps) as compound purified.

[0690] Using similar procedure for the synthesis of 343-YW-276 from343-YW-203, MK-049 (15.57 g, 74.05 mmol) was converted to MK-050 (19.51g, 72% 2 steps) as compound purified.

[0691] To a stirred solution of MK-050 (5.15 g, 14.16 mmol) in 35 mL ofdry THF was added n-BuLi in hexane (1.6M, 19.5 mL, 31.14 mmol) at −78°C. After 1 hr the reaction mixture was quenched with saturated aqueoussolution of NH₄Cl and diluted with AcOEt. The organic extract was washedwith brine, dried over MgSO₄, filtered and concentrated to give crudeproduct. It was purified by chromatography on silica gel (hexane/AcOEt:6/1 to 5/1) to afford oil of MK-051 (2.69 g, 93%).

[0692] Using similar procedure for the synthesis of NY-01 from TM-03,TM-03 (2.79 g, assumed to contain 10.0 mmol) coupled with MK-051 (2.68g, 13.1 mmol) was converted to crude alcohol. The crude product waspurified by chromatography on silica gel (hexane/AcOEt: 5/1 to 3/1) toafford oil of MK-052 (less polar single isomer, 986 mg, 20%) and oil ofMK-053

[0693] Using similar procedure for the synthesis of TM-05 from TM-04,MK-052 (less polar single isomer, 968 mg, 1.96 mmol) was converted tocolorless oil of MK-054 and purified (single isomer, 870 mg, 90%).

[0694] Using similar procedure for the synthesis of MK-024 from MK-023,MK-054 (834 mg, 1.69 mmol) treated with 3-MPMCl (0.61 mL, 4.21 mmol) wasconverted to colorless oil of MK-055 and purified (984 mg, 95%).

[0695] To a stirred solution of MK-055 (998 mg, 1.62 mmol) in 16 mL ofTHF was added TBAF in THF (1M, 2.43 mL, 2.43 mmol) at rt. After 3 hrsthe mixture was worked up in usual manner and purified by chromatographyon silica gel (hexane/AcOEt: 3/1) to afford colorless oil of MK-056 (696mg, 86%).

[0696] Using similar procedure for the synthesis of MK-008 from MK-007,MK-056 (695 mg, 1.39 mmol) was converted to crude aldehyde of MK-057(728 mg). The crude aldehyde was used for next step withoutpurification.

[0697] Using similar procedure for the synthesis of MK-044 from MK-043,MK-057 (728 mg, assumed to contain 1.39 mmol) was converted to colorlessoil of MK-058 and purified(643 mg, 90% 2 steps).

[0698] To a solution of MK-058 (200 mg, 0.389 mmol) and2,6-(^(t)Bu)₂-4-Me-pyridine (798 mg, 10 eq., 3.89 mmol) in 5 mL of dryCH₂Cl₂ was added (COCl)₂ in CH₂Cl₂ (2M, 0.97 ml, 5 eq., 1.94 mmol) at 0°C. and the solution was allowed to warm to rt. After stirred for 45 minthe reaction mixture was concentrated in vacuo under nitrogen atmosphereto give crude product

[0699] The crude product including MK-059 (assumed to contain 0.389 mmolderived from 0.389 mmol of MK-058, 1.03 eq.) was dissolved in 4 mL ofdry CH₂Cl₂ at 0° C. A solution of MK-048 (124 mg, 0.377 mmol) in 4 mL oftoluene was added and the mixture was allowed to warm to rt. Afterstirred for 15 min the reaction mixture was quenched with saturatedaqueous solution of NaHCO₃ and extracted with AcOEt. The organic extractwas washed with brine, dried over Na₂SO₄, filtered and concentrated togive crude product. It was purified by chromatography on silica gel(hexane/AcOEt: 3/1) to afford pale brown oil of MK-060 (297 mg,

[0700] To a stirred mixture of MK-060 (194 mg, 0.235 mmol) in 4 mL ofCH₂Cl₂ and 0.2 mL of aqueous phosphate buffer (pH 6.86) was added DDQ(59 mg, 1.1 eq., 0.259 mmol) at 0° C. After stirred for 1.5 hrs at 0° C.the reaction mixture was quenched with aqueous solution of NaHCO₃ andextracted with AcOEt. The organic extract was washed with saturatedaqueous solution of NaHCO₃ and brine, dried over Na₂SO₄, filtered andconcentrated to give crude oil. The crude product was purified bychromatography on silica gel (hexane/AcOEt: 5/2) to afford colorless oilof MK-061 (138 mg, 83%).

[0701] Using similar procedure for the synthesis of MK-031 from MK-030,MK-061 (165 mg, 0.234 mmol) was converted to crude MK-062 (159mg, >100%). The crude MK-062 was used for next step withoutpurification.

[0702] Ph₃P (221 mg, 0.844 mmol, 3.6 eq.) was dissolved in 39 mL of dryTHF and cooled to 0° C. 40% DEAD in toluene (0.32 mL, 0.703 mmol, 3.0eq.) was added and stirred for 20 min at 0° C. To the stirred solutionwas added dropwise over 15 min a solution of crude MK-062 (159 mg,assumed to contain 0.234 mmol) in 39 mL of THF at 0° C. After 10 min at0° C. the reaction mixture was evaporated and purified by chromatographyon silica gel (hexane/AcOEt: 2/1 to 3/2) to afford oil of MK-063 (115mg, 84% 2 steps).

[0703] To a stirred mixture of MK-063 (115 mg, 0.196 mmol) in 5 mL ofCH₂Cl₂ and 0.5 mL of aqueous phosphate buffer (pH 6.86) was added DDQ(103 mg, 2.3 eq., 0.452 mmol) at 0° C. and the mixture was allowed towarm to rt. After stirred for 24 hrs at rt the reaction mixture wasworked up in the usual manner to give a crude oil. The crude product waspurified by chromatography on silica gel (hexane/AcOEt: 2/1 to 1/3) toafford colorless oil of MK-064 (2^(nd) elution, 14 mg, 15%), colorlessoil of MK-065 (3^(rd) elution, 21 mg, 23%), and colorless oil of MK-066(1^(st) elution, 14 mg, 15%).

[0704] Using similar procedure for the synthesis of NF0552 from MK-021,MK-065 (21 mg, 0.0451 mmol) treated at low temperature was converted tocolorless oil of MK-066 (15 mg, 72%) as compound purified.

[0705] Using similar procedure for the synthesis of NF1226 from MK-035,MK-066 (29 mg, 0.0626 mmol) was converted to crude pale yellow crystals.The crude product was purified by chromatography on silica gel(hexane/AcOEt=1/3 to AcOEt alone) to afford colorless crystals of NF1535(17.6 mg, 74%).

[0706] Using similar procedure for the synthesis of NF1226 from MK-035,MK-064 (14 mg, 0.0302 mmol) was converted to crude pale yellow crystals.The crude product was purified by chromatography on silica gel(CH₂Cl₂/AcOEt=4/1, 2/1, to 1/1) to afford colorless crystals of NF1537(8.2 mg, 72%).

[0707] Synthetic Procedure for NF2306

[0708] To a stirred solution of methyl(R)-(−)-3-hydroxy-2-methylpropionate (7.00 g, 59.26 mmol) in 66 mL ofCH₂Cl₂ and 132 mL of cyclohexane were added CCl₃C(═NH)OBn (13.2 mL, 71.1mmol) and CF₃SO₃H (cat. 0.2 mL) at rt. After 3 hrs the reaction mixturewas diluted with hexane to form precipitation. After filteringprecipitation off, filtrates was washed with saturated aqueous NaHCO₃and brine, dried over Na₂SO₄, filtered, and concentrated to give a crudeoil. The crude product was purified by chromatography on silica gel(hexane/AcOEt=20/1) to afford colorless oil of MK-067 (9.60 g, 78%).

[0709] LiAlH₄ (2.62 g, 69.1 mmol) was suspended in 250 mL of dry THF. Tothe suspension was dropwise added a solution of MK-067 (9.59 g, 46.0mmol) in 57 mL of dry THF at 0° C. After stirred for 1 hr at 0° C. thereaction mixture was quenched with MeOH (13 mL), water (2.5 mL), 10%NaOH (2.5 mL), then water (7.5 mL). The mixture was dried over MgSO₄,filtered and evaporated to a crude product. The crude product waspurified by chromatography on silica gel (hexane/AcOEt=2/1) to affordcolorless oil of MK-068 (7.89 g, 95%).

[0710] Using similar procedure for the synthesis of MK-008 from MK-007,MK-068 (7.89 g, 43.76 mmol) was converted to crude MK-069 (8.66g,>100%). The crude MK-069 was used for next step without purification.

[0711] To a stirred suspension of CuI (10.83 g, 56.9 mmol, 1.3 eq.) in100 mL of dry Et₂O was added over 15 min MeLi in Et₂O (1.14 M, 98.6 mL,112.5 mmol, 2.57 eq.) at 0° C. After stirred for 30 min at 0° C., themixture was cooled to −78° C. Crude MK-069 (8.66 g, assumed to contain43.76 mmol) in 75 mL of dry Et₂O was added over 40 min at −78° C. thenthe mixture was stirred at −78° C. for additional 1 hr. The reactionmixture was allowed to warm to −20° C. over 1.5 hr, then quenched with28% aqueous NH₃ solution and extracted with AcOEt. The organic extractwas washed with brine, dried over MgSO₄, filtered and concentrated togive a crude oil. The crude product was purified by chromatography onsilica gel (hexane/AcOEt: 5/1) to afford pale yellow oil of MK-070 (6.89g, 81% 2 steps).

[0712] Ph₃P (9.24 g, 35.24 mmol, 1.4 eq.) was dissolved in 70 mL of dryTHF and cooled to 0° C. 40% DEAD in toluene (14.84 mL, 32.72 mmol, 1.3eq.) was added and stirred for 20 min at 0° C. To the stirred solutionwas dropwise added a solution of MK-070 (4.89 g, 25.17 mmol) and PhCO₂H(4.00 g, 32.7 mmol, 1.3 eq.) in 30 mL of dry THF at 0° C. After 30 minat 0° C. the mixture was allowed to warm to rt overnight. The resultingreaction mixture was evaporated and diluted with hexane-AcOEt. Afterfiltration of generated precipitate, the filtrate was concentrated toyield crude oil. The crude product was purified by chromatography onsilica gel (hexane/AcOEt: 15/1) to afford pale yellow oil of MK-071(6.84 g, 91%).

[0713] To a stirred solution of MK-071 (6.84 g, 22.91 mmol) in 38 mL ofEtOH was added aqueous 3N-NaOH (15.3 mL, 45.81 mmol) then the mixturewas stirred at 80° C. for 1 hr. The resulting mixture was evaporated,extracted with Et₂O, washed with brine, dried over MgSO₄, filtered andconcentrated to yield a crude oil. The crude product was purified bychromatography on silica gel (hexane/AcOEt: 5/1 to 5/3) to affordcolorless oil of MK-072 (4.17 g, 94%).

[0714] To a suspension of 66% NaH (916 mg, 25.20 mmol) in 30 mL of DMFwas added a solution of MK-072 (2.72 g, 14.00 mmol) in 10 mL of DMF at0° C. After stirring at 0° C. for 30 min 4-MPMCl (3.80 mL, 28.00 mmol)was added, then the mixture was allowed to warm to rt. After 2 days thereaction was quenched with saturated aqueous solution of NH₄Cl andextracted with AcOEt. The organic extract was washed with saturatedaqueous solution of NaHCO₃ and brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified by chromatography on silicagel (Hexane/AcOEt: 12/1) to afford colorless oil of MK-073 (3.88 g,88%).

[0715] 50 wt % suspension of Raney-Ni (W2) in basic water (9.5 g) wasadded into a flask, then the suspension was washed with water and EtOH.To this suspension was added a solution of MK-073 (3.88 g, 12.34 mmol)in 150 mL of EtOH. H₂ ballon was mounted and the mixture was purged withH₂. After stirred for 5 days under H₂ (1 atm) at rt, the reactionmixture was filtered through Celite and the filtrate was evaporated togive a crude oil. The crude product was purified by chromatography onsilica gel (Hexane/AcOEt: 3/1) to afford colorless oil of MK-074 (2.61g, 94%).

[0716] Using similar procedure for the synthesis of MK-008 from MK-007,MK-074 (2.61 g, 11.63 mmol) was converted to crude oil of MK-075 (2.70g, quant.). The crude MK-075 was used for next step withoutpurification.

[0717] Using similar procedure for the synthesis of 343-YW-276 from343-YW-203, crude MK-075 (2.70 g, assumed to contain 11.63 mmol) wasconverted to MK-076 (3.92 g, 89% 2 steps).

[0718] MK-076 (2.03 g, 5.37 mmol, 1.36 eq.) was dissolved in 27 mL ofdry THF and cooled to −78° C. under nitrogen. n-BuLi in hexane (1.6M,6.71 mL, 10.73 mmol, 2.71 eq.) was added and stirred at −78° C. for 1hr. A solution of crude TM-03 (1.11 g, assumed to contain 3.96 mmol) in7 mL of dry THF was dropwise added to the mixture and stirred for 30 minat −78° C. The reaction mixture was allowed to warm to 10° C. slowlyover 2.5 hrs. The mixture was quenched with saturated aqueous solutionof NH₄Cl and extracted with AcOEt. The organic extract was washed withsaturated aqueous solution of NaHCO₃ and brine, dried over Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel (hexane/AcOEt: 7/1 to 4/1) to afford oil ofMK-077 (less polar single isomer, 541 mg, 27%) and oil of MK-078 (polarsingle isomer, 1.21 g, 60%).

[0719] Using similar procedure for the synthesis of TM-05 from TM-04,MK-077 (less polar single isomer, 3.85 g, 7.61 mmol) was converted tocolorless oil of MK-079 (single isomer, 3.38 g, 87%).

[0720] Using similar procedure for the synthesis of MK-024 from MK-023,MK-079 (3.38 g, 6.63 mmol) treated with 3-MPMCl (2.89 mL, 19.9 mmol) wasconverted to colorless oil of MK-080 (3.86 g, 92%).

[0721] Using similar procedure for the synthesis of MK-056 from MK-055,MK-080 (3.85 g, 6.12 mmol) was converted to colorless oil of MK-081(3.00 g, 95%).

[0722] Using similar procedure for the synthesis of MK-058 from MK-056,MK-081 (1.82 g, 3.54 mmol) was converted to colorless oil of MK-083(1.80 g, 96% 2 steps).

[0723] Using similar procedure for the synthesis of MK-059 from MK-058,MK-083 (1.80 g, 3.40 mmol) was converted to crude oil of MK-084. Thecrude MK-084 was used for next step without purification.

[0724] Using similar procedure for the synthesis of MK-060 from MK-059,crude MK-084 coupled with MK-048 (853 mg, 2.60 mmol) was converted topale brown oil of MK-085 (2.14 g, 98% 2 steps).

[0725] Using similar procedure for the synthesis of MK-061 from MK-060,MK-085 (2.14 g, 2.55 mmol) was converted to pale yellow oil of MK-086(1.74 g, 95%).

[0726] Using similar procedure for the synthesis of MK-031 from MK-030,MK-086 (1.74 g, 2.42 mmol) was converted to crude oil of MK-087 (1.57 g,quant.). The crude MK-087 was used for next step without purification.

[0727] Using similar procedure for the synthesis of MK-063 from MK-062,crude MK-087 (1.57 g, assumed to contain 2.42 mmol) was converted topale yellow oil of MK-088 (1.68 g, including ca 0.38 g of inseparableimpurity derived from DEAD, ca 90% 2steps).

[0728] Using similar procedure for the synthesis of MK-065 from MK-063,MK-088 (1.68 g, including ca 0.38 g of inseparable impurity derived fromDEAD, assumed to contain 2.17 mmol) was converted to colorless solid ofMK-089 (525 mg, 50%).

[0729] Using similar procedure for the synthesis of NF0552 from MK-021,MK-089 (458 mg, 0.955 mmol) was treated with Dess-Martin reagent at lowtemperature to give a pale yellow solid of MK-090 (250 mg, 55%).

[0730] MK-090 (250 mg, 0.524 mmol) was dissolved in 3.5 mL of CH₂Cl₂ andcooled to 0° C. A mixture of 50% Hydrofluoric acid (24N, 3.5 mL) and 14mL of CH₃CN was added to the solution and stirred for 1 hr at 0° C.before the mixture was allowed to warm to 15° C. slowly over 1.5 hrs.Then, the reaction mixture was poured into a stirred biphasic solutionof saturated aqueous solution of NaHCO₃ and AcOEt. The organic extractwas washed with brine, dried over Na₂SO₄, filtered and concentrated tocrude product. The crude product was purified by chromatography onsilica gel (CH₂C₂/MeOH: 13/1) to afford colorless crystals of NF2306(174 mg, 42%).

[0731] Preparation of C13-Oxygen and Fluoro Analog, NF2432, NF2544,NF2547, NF2553 and NF2556

[0732] Synthetic Procedure for NF2432

[0733] 1) Preparation of the Macrocyclic Part

[0734] To a stirred THF solution of dibromide YE-43 (18.6 g, 37 mmol),which was prepared using similar procedure for the intermediate554-RB-228 from methyl (S)-3-hydroxybutyrate (47%, 5 steps), was addedn-butyllithium (1.6M in hexane solution, 47 ml, 75 mmol) at −78° C. Themixture was allowed to warm to 0° C. for 30 min and then stirred at −78°C. for additional 30 min. A THF solution of aldehyde NY-20 (6.17 g, 24mmol) was added, the mixture was allowed to warm to 0° C. and stirred at0° C. for 30 min, after which a saturated solution of NH₄Cl was added.The mixture was extracted with EtOAc and the organic extract was washedwith a saturated solution of NaHCO₃, brine, dried over anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 12% EtOAc/hexane to give 3.55 g (5.9mmol, 25%) of the less polar isomer YE-01 and 5.5 g (0.093 mol, 39%) ofthe more polar isomer YE-02.

[0735] To a solution of YE-02 (10.1 g, 17.0 mmol) in n-hexane (170 mL),quinoline (0.25 eq., 4.25 mmol, 0.50 mL) and 5 wt. % Pd on BaSO₄ (0.05eq., 0.85 mmol, 1.81 g) was added at rt. And the reaction mixture waspurged with H₂ and stirred under H₂ atmosphere for 11 hrs. The catalystwas filtered off and the filtrate was concentrated. The crude product(11.5 g) was purified by chromatography on silica gel using 16%EtOAc/hexane to give 8.19 g (13.7 mmol, 81% 2 steps) of the desiredallylic alcohol YE-03.

[0736] The alcohol YE-03 (8.04 g, 13.5 mmol) was dissolved in CH₂Cl₂(200 mL), 2,6-lutidine (7.8 mL, 67.0 mmol) was added and the mixture wascooled to 0° C. in ice/water bath. Then TBSOTf (7.7 mL, 33.5 mmol) wasadded and the mixture was allowed to warm to rt. After 2 hrs, it wascooled to 0° C. and was quenched with MeOH and a saturated solution ofNaHCO₃. The mixture was extracted with EtOAc, the organic layer waswashed with a saturated solution of NaHCO₃, 5% citric acid aq., asaturated solution of NaHCO₃, and brine. The organic layer was driedover Na₂SO₄, filtered and concentrated. The crude product was purifiedby chromatography on silica gel using 2-4% EtOAc/hexane to give 9.60 g(13.5 mmol, quant.) of YE-04.

[0737] To a stirred ether solution (200 ml) of YE-04 (9.60 g, 13.5 mmol)at 0° C. was added lithium tetrahydroborate (0.60 g, 27.5 mmol). Themixture was allowed to warm to rt and stirred for 2 days. Then themixture was cooled to 0° C. and lithium tetrahydroborate (0.30 g, 13.8mmol) was added again. The mixture was allowed to warm to rt and stirredovernight. The mixture was cooled to 0° C., then a saturated solution ofNH₄Cl (2ml) was added slowly. After the stirring for 20 min, a saturatedsolution of NH₄Cl (100 ml) was added. The mixture was extracted withEtOAc and the organic extract was washed with a saturated solution ofNH₄Cl, brine, dried over anhydrous Na₂SO₄, filtered and concentrated.The crude product was purified by chromatography on silica gel using12-15% EtOAc/hexane to give 8.51 g (13.5 mmol, quant.) of the alcoholYE-05.

[0738] To a stirred solution of YE-05 (1.90 g, 3.03 mmol) in toluene (45ml) was added triphenylphosphine (1.6 g, 6.1 mmol), a mixture of diethylazodicarboxylate (40% in toluene, 2.1 ml, 4.6 mmol) and iodomethane(0.29 ml, 4.7 mmol). The mixture was stirred for 40 min after which amixture of diethyl azodicarboxylate (in toluene, 0.5 ml, 1.1 mmol) andiodomethane (0.06 ml, 1.2 mmol) was added. The mixture was stirred for20 min and the solvent was evaporated in vacuo. The concentrate waspurified by chromatography on silica gel using 1-1.5% EtOAc/hexane togive 2.05 g (2.78 mmol, 92%) of the iodide YE-06.

[0739] 2) Preparation of the Aromatic Part

[0740] 3-Bromo-4-hydroxy-5-methoxybenzaldehyde (24.8 g, 0.107 mol) wasdissolved in DMF (400 ml). K₂CO₃ (20 g, 0.14 mol) and iodomethane (8.8ml, 0.14 mol) was added. The mixture was stirred for 4 hrs and was thencooled to 0° C. in ice/water bath and diluted with ether (300 ml). Thenice-water (600 ml) was added slowly. The mixture was extracted withether and the organic extract was washed with water, brine, dried overanhydrous Na₂SO₄, filtered and concentrated to give 22.1 g (90 mmol,84%.) of YE-07.

[0741] YE-07 (13.3 g, 54.2 mmol) was dissolved in chloroform (350 mL)under nitrogen. m-CPBA (>70%, 31 g, 126 mmol) was added and the solutionwas gently refluxed for 1 hr. The reaction mixture was cooled to 0° C.and was poured into saturated stirred solution of NaHCO₃. After stirringfor 15 min, the organic layer was separated, washed with saturatedNa₂SO₃, saturated NaHCO₃, dried over Na₂SO₄, filtered and concentrated

[0742] The concentrate was dissolved in MeOH (150 ml) and 6N HCl (150ml) was added. The mixture was stirred for 15 min. and partiallyconcentrated. The mixture was extracted with EtOAc and the organicextract was washed with water, dried over Na₂SO₄, filtered andconcentrated. to give 10.8 g of crude phenol.

[0743] The crude phenol was methylated using the same procedure as thepreparation of YE-07 to give 9.8 g (39.7 mmol, 73%) of YE-08.

[0744] To a stirred ether solution of bromobenzene YE-08 (14.6 g, 59.1mmol) at −78° C. was added n-butyllithium (1.6M in hexane solution, 50ml, 1.3 eq., 80 mmol,), and the mixture was stirred at −78° C. for 1.5hrs. A solution of iodomethane (10 ml, 161 mmol) in ether (20 ml) wasadded, and the mixture was allowed to warm to rt and stirred for 1.5hrs, after which a saturated solution of NH₄Cl was added. The mixturewas extracted with ether and the organic extract was washed with brine,dried over anhydrous Na₂SO₄, filtered and concentrated. The concentratewas purified by chromatography on silica gel using 6-9% EtOAc/hexane togive 9.6 g (52.6 mmol, 89%) of 2,3,5-trimethoxytoluene.

[0745] The toluene 9.6 g (52.6 mmol, 89%) was dissolved in DME (130 mL)and copper (II) bromide (25 g, 112 mmol) was added portionwise over 6hrs. After stirring another 1 hr, the reaction mixture was filtrated andthe filtrate was concentrated. The concentrate was purified bychromatography on silica gel using 5-12% EtOAc/hexane to give 12.7 g(48.6 mmol, 92%) of YE-09.

[0746] To a stirred ether solution of YE-09 (45.6 mmol, 11.9 g) at −78°C. was added n-butyllithium (1.6M in hexane solution, 37 ml, 59 mmol)and the mixture was stirred at −78° C. for 1 hr. A finely crusheddry-ice was added slowly and the mixture was allowed to warm to −10° C.After 2 hrs, the reaction was quenched with water (200 ml). The mixturewas washed with ether then acidified with 1N HCl. The mixture wasextracted with EtOAc and the organic extract was washed with water,brine, dried over anhydrous Na₂SO₄, filtered and concentrated to give9.7 g (42.8 mmol, 94%) of YE-10.

[0747] To a stirred solution of YE-10 (1.4 g, 6.3 mmol) in dry CH₂Cl₂(40 mL) was added BBr₃ (1M solution, 28 mL, 28 mmol) at −78° C. undernitrogen atmosphere and the mixture was allowed to warm to rt. After 8hrs, the mixture was cooled to 0° C. and poured into water. The organiclayer was separated and washed with a solution of 5% glycerol, brine,dried over anhydrous Na₂SO₄, filtered and concentrated to give 0.9 g ofthe crude product.

[0748] To a stirred solution of the crude product (0.9 g, 4.8 mmol) indry acetonitrile (20 mL) were added MeOH (0.78 ml) followed byN,N-diisopropylethylamine (1.7 ml, 9.7 mmol). To the mixture was addedTMSCHN₂ (2.0M in hexane, 4.8 mL, 9.6 mmol) and the mixture was stirredat 30° C. After 1 hr., the mixture was cooled to 0° C., poured intowater and extracted with EtOAc. The organic extract was washed with asaturated solution of NH₄Cl, water, brine, dried with anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 6% EtOAc/hexane to give 0.37 g (1.74mmol, 28% 2steps) of YE-11.

[0749] To a stirred suspension of NaH (0.16 g, 4.4 mmol) in dry THF (10mL) was added YE-11 (0.37 g, 1.7 mmol) in dry THF (8 mL) at 0° C. Afterstirring for 30 min at 0° C., TBDPSCl (0.5 ml, 1.9 mmol) was added. Themixture was allowed to warm to rt and stirred for 15 min. The mixturewas cooled to 0° C., poured into water and extracted with EtOAc. Theorganic extract was washed with water, brine, dried with anhydrousNa₂SO₄, filtered and concentrated. The concentrate was purified bychromatography on silica gel using 6% EtOAc/hexane to give 0.63 g (1.39mmol, 80%) of silyl ether.

[0750] To a stirred solution of the silyl ether (3.7 g, 8.3 mmol) in DMF(50 mL) was added Cs₂CO₃ (3.0 g, 9.2 mmol) and iodomethane (1.3 mL, 20.8mmol). The mixture was stirred overnight and was cooled to 0° C. inice/water bath. Then the mixture was poured into ice-cold, saturatedsolution of NH₄Cl (100 mL) and extracted with EtOAc. The organic extractwas washed with water, brine, dried over anhydrous Na₂SO₄, filtered andconcentrated. The concentrate was purified by chromatography on silicagel using 5% EtOAc/hexane to give 2.83 g (6.09 mmol, 73%) of YE-12.

[0751] Using similar procedure for the intermediate 10 of NF2561, YE-12(0.30 g, 0.64 mmol) was converted to YE-13 (0.29 g, 0.51 mmol, 80% 2steps).

[0752] YE-13 (0.29 g, 0.51 mmol) was dissolved in THF (10 mL). Then 1.0Msolution of TBAF in THF (1.5 mL, 1.5 mmol) were added at rt. The mixturewas stirred overnight after which a saturated solution of NH₄Cl wasadded. The mixture was extracted with EtOAc and the organic extract waswashed with a saturated solution of NaHCO₃, water, brine, dried withanhydrous Na₂SO₄, filtered and concentrated. The crude product waspurified by chromatography on silica gel using 10% EtOAc/hexane to EtOAcas eluents to give 0.17 g (0.51 mmol, quant.) of phenol.

[0753] To a stirred suspension of sodium hydride (0.28 g, 7.7 mmol) indry THF(10 mL) was added the phenol (2.0 g, 5.9 mmol) in dry THF (20mL)at 0° C. After stirring for 15 min, chloromethyl methyl ether (0.57mL, 7.5 mmol) at 0° C. After 3 hr, the mixture was poured into asaturated solution of NH₄Cl and extracted with EtOAc. The organicextract was washed with a saturated solution of NaHCO₃, brine, driedover anhydrous Na₂SO₄, filtered and concentrated. The crude product waspurified by chromatography on silica gel using 15% EtOAc/hexane to give1.4 g (3.7 mmol, 62%) of YE-14.

[0754] Using similar procedure for the intermediate 14 of NF2561, YE-14(1.4 g, 3.7 mmol) was converted to YE-15 (1.6 g, 3.4 mmol, 93% 2steps).

[0755] Using similar procedure for the intermediate 18 of NF2561, theiodide (502 mg, 0.68 mmol) was converted to YE-16 (315 mg, 0.33 mmol,48% 3 steps).

[0756] YE-16 (310 mg, 0.32 mmol) was dissolved in THF (12 mL). Then,1.0M solution of tetrabutylammonium fluoride in THF (1.6 mL, 1.6 mmol)was added at 0° C. The mixture was stirred at rt for 2 days after which10% KHSO₄ solution was added. The mixture was extracted with EtOAc andthe organic extract was washed with water, brine, dried with anhydrousNa₂SO₄, filtered and concentrated. The crude product was dried byazeotropic distillation with toluene to give 270 mg of YE-17 with silylimpurity.

[0757] To a stirred solution of crude YE-17 (270 mg) in THF (20 mL) wereadded triethylamine (0.090 mL, 0.64 mmol) and 2,4,6-trichlorobenzoylchloride (0.085 mL, 0.54 mmol) at rt. After 16 hrs, the reaction mixturewas diluted with toluene (300 mL) and added dropwise to a solution of4-(dimethylamino)pyridine (980 mg, 8.0 mmol) in toluene (320 mL) over aperiod of 6 hrs under reflux. The resultant mixture was stirred for 0.5hr under reflux. After concentration under reduced pressure, the residuewas dissolved in EtOAc and washed with 10% KHSO₄ aq sol., water, brineand dried over anhydrous Na₂SO₄, filtered and concentrated. The crudeproduct was purified by chromatography on silica gel using 6-30%EtOAc/hexane to give 114 mg (0.23 mmol, 72% 3steps) of YE-18.

[0758] Using similar procedure for 509-HD-125, YE-18 (20 mg, 0.041 mmol)was converted to YE-19 (22 mg, quant.).

[0759] To a stirred solution of YE-19 (22 mg, 0.041 mmol) in THF (1.4mL)-H₂O (0.7 mL) was added trifluoroacetic acid (1.4 mL) at 0° C. Themixture was then allowed to warm to rt. After 1.5 hrs, the mixture waspoured into a saturated solution of NaHCO₃ and extracted with EtOAc. Theorganic extract was washed with water, brine and dried over anhydrousNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel using 30% EtOAc/hexane to give 13.3 mg(0.033 mmol, 81%) of NF2432.

[0760] Synthetic Procedure for NF2544

[0761] To a stirred suspension of NaH (2.3 g, 63 mmol) in dry THF (100mL) were added 2-bromo-4-fluorophenol (10 g, 52 mmol) in dry THF (20 mL)at 0° C. After stirring for 30 min at 0° C., chloromethyl methyl ether(4.8 mL, 63 mmol) was added. The mixture was allowed to warm to rt andstirred for 1.5 hrs. The mixture was cooled to 0° C., poured into waterand extracted with EtOAc. The organic extract was washed with water,brine, dried with anhydrous Na₂SO₄, filtered and concentrated. Theconcentrate was purified by chromatography on silica gel using 10%EtOAc/hexane to give 10.7 g (45.5 mmol, 87%) of YE-20.

[0762] To a stirred suspension of YE-20 (10.7 g, 46 mmol) in dry ether(150 mL) was added 1.6M n-BuLi in hexane (34 mL, 54.4 mmol) at −78° C.under nitrogen atmosphere. After 1 hr, dry DMF (15 mL) was added and themixture was allowed to warm to rt. It was quenched with a saturatedsolution of NH₄Cl and extracted with EtOAc. The organic extract waswashed with a saturated solution of NH₄Cl, water, brine, dried overanhydrous Na₂SO₄, filtered and concentrated. The crude product waspurified by chromatography on silica gel using 10% EtOAc/hexane to give7.8 g (42 mmol, 93%) of YE-21.

[0763] To a stirred solution of YE-21 (6.8 g, 37 mmol) in toluene (200mL) was added ethylene glycol (12 g, 193 mmol) and p-toluenesulfonicacid monohydrate (0.3 g) at rt. and refluxed using Dean-Stark apparatus.After 4 hr, the mixture was cooled to 0° C. in ice/water bath. Then TEA(15 ml, 0.10 mol) was added, the mixture was stirred for 10 min, thenpoured into a saturated solution of NaHCO₃. The mixture was extractedwith EtOAc and the organic extract was washed with a saturated solutionof NaHCO₃, brine, dried over anhydrous Na₂SO₄, filtered and concentratedto give 6.7 g of phenol as crude product.

[0764] To a stirred suspension of NaH (1.5 g, 41 mmol) in dry THF (120mL) were added the phenol (6.7 g, 36 mmol) in dry THF (20 mL) at 0° C.After stirring for 10 min at 0° C., TBSCl (6.3 g, 42 mmol) was added.The mixture was allowed to warm to rt and stirred overnight. The mixturewas cooled to 0° C., poured into water and extracted with EtOAc. Theorganic extract was washed with water, brine, dried over anhydrousNa₂SO₄, filtered and concentrated. The concentrate was purified bychromatography on silica gel using 2% EtOAc/hexane (containing 0.2% oftriethylamine) to give 8.75 g (29.3 mmol, 79% 2steps) of YE-22.

[0765] To a stirred suspension of YE-22 (20 g, 67 mmol) in THF (350 mL)was added 1.6M n-BuLi in hexane (50 mL, 80 mmol) at −78° C. undernitrogen atmosphere. After 1 hr TMEDA(15 mL, 99 mmol) was added andstirred at −78° C. for another 10 min. To the mixture was added dry DMF(5.0 mL, 65 mmol) and the mixture was allowed to warm to 31 20° C. andstirred for 40 min. It was quenched with AcOH (14 mL), and poured intowater. The mixture was extracted with EtOAc and the organic extract waswashed with a saturated solution of NaHCO₃, brine, dried over anhydrousNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel using 2-5% EtOAc/hexane to give 15.8 g(48.4 mmol, 72%) of YE-23.

[0766] To a stirred MeOH solution (350 mL) of YE-23 (15.8 g, 48.4 mmol)at 0° C. was added NaBH₄ (2.0 g, 53 mmol). The mixture was stirred at 0°C. for 30 min. Then the mixture was quenched with AcOH (10 mL), andpoured into a saturated solution of NaHCO₃. The mixture was extractedwith ether and the organic extract was washed with a saturated solutionof NaHCO₃, brine, dried over anhydrous Na₂SO₄, filtered and concentratedto give 16.4 g (48.4 mmol, quant.) of alcohol.

[0767] To a stirred acetone solution (300 mL) of the alcohol (16.4 g,48.4 mmol) at 0° C. was added 1N HCl (8 mL, 8 mmol). The mixture wasstirred at 0° C. for 70 min, then was poured into a saturated solutionof NaHCO₃. The mixture was partially concentrated to remove acetone andthe concentrate was extracted with ether. The organic extract was washedwith brine, dried over anhydrous Na₂SO₄, filtered and concentrated togive 13.6 g (47.8 mmol) of YE-24.

[0768] YE-24 (13.6 g, 47.8 mmol) was dissolved in 900 mL ofdichloromethane and PDC (53.7 g, 143 mmol) were added to the solution.The reaction mixture was stirred for 7 d at rt and the mixture wasdiluted with ether (300 mL). After passing through Florisil column,YE-25 was obtained as colorless oil (11.7 g, 86%, 3steps).

[0769] YE-25 (11.7 g, 41.4 mmol) was dissolved in THF (12 mL), 1.7Nhydrochloric acid (60 mL, 100 mmol) were added and the mixture wasstirred at 50° C. for 1 hrs, then refluxed for 3 hrs. The mixture wascooled to rt and filtrated. To the filtrated was added 5N NaOH (18 mL)and the organic layer was separated. The aqueous layer was saturatedwith NaCl and was extracted with EtOAc. The combined organic extaractwas concentrated and the concentrate was triturated with ether to givephenol (4.9 g, 32.2 mmol).

[0770] Using similar procedure for TM-37, the phenol (4.2 g, 27.6 mmol)was converted to YE-26 (4.2 g, 21.2 mmol, 77%).

[0771] Using similar procedure for TM-50, YE-26 (4.1 g, 20.9 mmol) wasconverted to YE-27 (4.9 g, 14.6 mmol, 70% 3 steps).

[0772] Using similar procedure for TM-39, YE-27 (1.5 g, 4.5 mmol) wasconverted to YE-28 (1.60 g, 3.8 mmol, 85% 2 steps).

[0773] Using similar procedure for the intermediate 18 of NF2561, theiodide (510 mg, 0.69 mmol) was converted to YE-29 (690 mg, 0.67 mmol,97% 3 steps).

[0774] Using similar procedure for the intermediate YE-17, YE-29 (520mg, 0.57 mmol) was converted to YE-30 (430 mg) with silyl impurity.

[0775] Using similar procedure for YE-18, YE-30 (430 mg) was convertedto YE-31 (94 mg, 0.21 mmol, 36.5% 3 steps).

[0776] Using similar procedure for ER803064, YE-31 (45 mg, 0.099 mmol)was converted to NF-2544 (13 mg, 0.036 mmol, 36% 2 steps).

[0777] Synthetic Procedure for NF2547

[0778] 5-Bromovanillin (20.0 g, 86.56 mmol) and AlCl₃ (12.70 g, 95.22mmol) were dissolved in 145 mL of dry CH₂Cl₂. To the stirred solutionwas dropwise added pyridine (30.80 mL, 380.9 mmol) over 10 min at rt(exothermic reaction). Then the mixture was warmed to 45° C. and stirredfor 20 hrs after which it was cooled to rt. The resulting mixture wasacidified with 3N HCl aq and extracted with AcOEt. The organic extractwas washed with brine, dried over Na₂SO₄, filtered and concentrated togive crude colorless crystals of MK-101 (18.4 g, <98%). The crude MK-101was used for next step without purification.

[0779] Crude MK-101 (13.15 g, assumed to contain 60.59 mmol), Br—CH₂—Cl(6.50 mL, 96.9 mmol), and Cs₂CO₃ (31.59 g, 96.9 mmol) were suspended in200 mL of DMF and the mixture was stirred at 110° C. for 20 hrs. Theresulting mixture was diluted with water and extracted with AcOEt. Theorganic extract was washed with brine, dried over Na₂SO₄, filtered andconcentrated to give crude brown solid of MK-102. The crude product waspurified by chromatography on silica gel (hexane/AcOEt: 6/1 to 5/1) toafford colorless crystals of MK-102 (11.1 g, 77% 2 steps).

[0780] 70% mCPBA (13.1 g, 53.1 mmol) was added to a stirred solution ofMK-102 (5.07 g, 22.12 mmol) in 110 mL of CHCl₃ and the mixture washeated up to reflux. After 3 hrs at reflux the mixture was cooled to 0°C., quenched with aqueous solution of Na₂SO₃, and extracted with CH₂Cl₂.The organic extract was washed with saturated aqueous solution of NaHCO₃and brine, then evaporated to give crude pale brown solid ofintermediate formate.

[0781] The crude formate was dissolved in 100 mL of MeOH. NaHCO₃ (3.7 g,44 mmol) was added and the suspension was stirred at rt for 30 min.Water and AcOEt were added to the mixture and organic extract was washedwith brine, dried over Na₂SO₄, filtered and concentrated to give a crudeoily solid of MK-103. The crude product was purified by chromatographyon silica gel (hexane/AcOEt: 4/1) to afford colorless crystals of MK-103(3.78 g, 79% 2 steps).

[0782] Using similar procedure for the synthesis of 509-HD-209 from509-HD-207, MK-103 (3.78 g, 17.40 mmol) was converted to colorless oilof MK-104 (4.42 g, 97%).

[0783] To a stirred solution of MK-104 (2.00 g, 7.66 mmol) in 26 mL ofdry Et₂O was dropwise added n-BuLi in hexane (1.6M, 5.74 mL, 9.19 mmol,1.2 eq.) at −78° C. After 1 hr dry DMF (1.20 mL, 15.3 mmol, 2.0 eq.) wasadded in one portion to the mixture and stirred at −78° C. for 30 minafter which saturated aqueous solution of NH₄Cl was added. The resultingmixture was extracted with AcOEt and the organic extract was washed withbrine, dried over Na₂SO₄, filtered, and concentrated to give crudeyellow oil of MK-105 (1.60 g). The crude product was used for next stepwithout purification.

[0784] The crude MK-105 (1.60 g, assumed to contain 7.66 mmol) wasdissolved in 26 mL of MeOH and the stirred solution was cooled to 0° C.NaBH₄ (290 mg, 7.66 mmol) was added in some portions. After 20 min at 0°C. the reaction mixture was quenched with aqueous solution of NH₄Cl andextracted with AcOEt. The organic extract was washed with brine, driedover Na₂SO₄, filtered and concentrated to give crude oil. The crudeproduct was purified by chromatography on silica gel (hexane/AcOEt: 3/1to 2/1) to afford colorless crystals of MK-106 (1.44 g, 89% 2 steps).

[0785] To a stirred solution of MK-106 (1.26 g, 5.92 mmol) in 30 mL ofDMF was added NBS (1.16 g, 6.51 mmol, 1.1 eq.) at 0° C. The mixture wasstirred at 0° C. for 3 hrs before aqueous solution of Na₂SO₃ and AcOEtwere added. The organic extract was washed with brine, dried overNa₂SO₄, filtered, and concentrated to give crude colorless crystals ofMK-107 (1.73 g, quant.). The crude product was used for next stepwithout purification.

[0786] Crude MK-107 (1.73 g, assumed to contain 6.51 mmol) and imidazole(1.01 g, 14.80 mmol) were dissolved in 39 mL of DMF. TBS-Cl (1.78 g,11.84 mmol) was added to the solution and the mixture was stirred at rtovernight after which AcOEt was added. The resulting mixture was washedwith aqueous solution of NaHCO₃ and brine, dried over Na₂SO₄, filteredand concentrated to give crude crystals. The crude product was purifiedby chromatography on silica gel (hexane/AcOEt: 9/1) to afford colorlesscrystals of MK-108 (2.25 g, 94% 2 steps).

[0787] MK-108 (2.25 g, 5.56 mmol) was suspended in 90 mL of dry Et₂O. Tothe stirred suspension was added n-BuLi in hexane (1.6M, 4.52 mL, 7.23mmol, 1.3 eq.) and the mixture was stirred at −78° C. After 1.5 hr thesuspension turned homogeneous. The mixture was stirred for additional 30min at −78° C. Then, excessive dry CO₂ gas (ca 30 eq.) was added bybubbling through an inlet over 15 min. The resulting mixture was stirredat −78° C. for 40 min after which it was allowed to warm to rt. After 1hr the reaction mixture was quenched with saturated aqueous solution ofNa₂CO₃ and washed with Et₂O. The basic aqueous layer was acidified withKHSO₄ and extracted with AcOEt. The organic extract was washed withbrine, dried over Na₂SO₄, filtered, and concentrated to yield crude paleyellow crystals of MK-109 (1.93 g). The crude product was used for nextstep without purification.

[0788] Using similar procedure for the synthesis of MK-093 from MK-092,crude MK-109 (1.93 g) was converted to a crude brown oil of MK-110 (1.73g). The crude product was used for next step without purification.

[0789] Crude MK-110 (1.73 g) was dissolved in 75 mL of THF. Then, TBAFin THF (1M, 9.0 mL, 9.0 mmol, 2 eq.) was added at rt. The mixture wasstirred for 2 days before AcOEt and aqueous solution of KHSO₄C wereadded. The organic extract was washed with brine, dried over Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel (hexane/AcOEt: 2/1) to afford colorlesscrystals of MK-111 (476 mg, 36% 4 steps).

[0790] Using similar procedure for the synthesis of NY-90 from NY-89,MK-111 (412 mg, 1.73 mmol) was converted to crude pale yellow oil ofMK-112 (435 mg). The crude product was used for next step withoutpurification.

[0791] Using similar procedure for the synthesis of NY-91 from NY-90,crude MK-112 (435 mg) was converted to colorless oil of MK-113 (494 mg,79% 3 steps).

[0792] To a stirred solution of MK-113 (423 mg, 1.17 mmol) in 9 mL ofDMSO was added a solution of KOH (196 mg, 3.50 mmol) in 4.5 mL of waterand the mixture was heated to 80° C. After stirred for 2 hrs, themixture was cooled to rt and diluted with Et₂O. The mixture wasacidified with aqueous solution of KHSO₄ and extracted with AcOEt. Theorganic extract was washed with brine, dried over Na₂SO₄, filtered, andconcentrated to give crude pale yellow crystals of MK-114 (428 mg). Itwas used for next step without purification.

[0793] Using similar procedure for the synthesis of MK-047 from MK-046,crude MK-114 (428 mg) was converted to pale pink oil of MK-115 (476 mg,91% 2 steps) as compound purified.

[0794] Using similar procedure for the synthesis of compound 18, theintermediate of NF2561, from compound 16, YE-06 (627 mg, 0.851 mmol)coupled with MK-115 (474 mg, 1.06 mmol) was converted to pale yellow oilof MK-116 (804 mg, 99% 3 steps).

[0795] Using similar procedure for the synthesis of YE-17 from YE-16,MK-116 (800 mg, 0.844 mmol) was converted to crude oil of MK-117 (735mg, including silyl impurity). The crude product was used for next stepwithout purification.

[0796] Using similar procedure for the synthesis of YE-18 from YE-17,the crude MK-117 (735 mg, assumed to contain 0.844 mmol) was convertedto a colorless solid of MK-118 (192 mg, 48% 3 steps).

[0797] Using similar procedure for the synthesis of 509-HD-125 from509-HD-119B, MK-118 (141 mg, 0.296 mmol) was converted to crude paleyellow solid of MK-119 (104 mg, <74%). The crude MK-119 was used fornext step without purification.

[0798] Using similar procedure for the synthesis of NF2306 from MK-090,crude MK-119 (104 mg) was converted to crude pale yellow oil. The crudeproduct was purified by chromatography on silica gel (hexane/AcOEt: 1/2)to afford pale yellow crystals of NF2547 (59 mg, 51% 2 steps).

[0799] Synthetic Procedure for NF-2553

[0800] Using same procedure for 509-HD-209, NY-37 (4.8 g, 19.59 mmol)was converted to NY-117 (4.93 g).

[0801] NY-117 (2.89 g, 10 mmol) was dissolved in Et₂O (40 mL) and cooledto −78° C., under nitrogen. Then, sec-BuLi (1.3M/cyclohexane, 9.3 mL,12.09 mmol) was slowly added and the reaction was stirred at −78° C. for30 min. DMF (1.55 mL, 20 mmol) was added to the solution, then thesolution was stirred at −78° C. for 10 min. The mixture was quenchedwith sat.NH₄Cl and extracted with EtOAc. The organic layer was washedwith water, brine and dried over Na₂SO₄, filtered and concentrated. Thecrude product was purified on silica gel column with hexane/EtOAc, 15:1,10:1, 8:1, 6:1, 5:1, 4:1 to give 811 mg of NY-118.

[0802] A mixture of NY-118 (4.09 g, 17.17 mmol), mCPBA (10 g, 40.56mmol) and CHCl₃ (15 mL) was refluxed for 2.5 hrs. The reaction mixturewas quenced with sat. Na₂S₂O₃ and extracted with EtOAc. The organiclayer was washed with sat. Na₂S₂O₃, sat.NaHCO₃(×2), brine, dried overNa₂SO₄, filtered and concentrated. The crude product was purified onsilica gel column with hexane/EtOAc, 10:1, 8:1 to give 2.58 g of NY-119.

[0803] Using same procedure for 10, NY-119 (2.58 g, 10.15 mmol) wasconverted to NY-120 (2.45 g).

[0804] A mixture of NY-120 (470 mg, 1.41 mmol), 2-bromo-1-chloroethane(0.35 ml, 4.22 mmol), K₂CO₃ (292 mg, 2.11 mmol) was refluxed for 6 hrs.Then, additional 2-bromo-1-chloroethane (0.35 ml 4.22 mmol) was addedand the mixture was refluxed for 15 hrs. K₂CO₃ (450 mg, 3.26 mmol) and2-bromo-1-chloroethane (1.05 ml, 12.67 mmol) were added and the mixturewas refluxed for 4 hrs. The insoluble material was filtered and thefiltrate was concentrated. The residue was diluted with EtOAc and washedwith water, brine, dried over Na₂SO₄, filtered and concentrated to give519 mg of NY-121.

[0805] A mixture of NY-121 (519 mg, 1.31 mmol), NaN₃ (212 mg, 3.26 mmol)and DMF (10 mL) was stirred at 80° C. for 3 hrs. The mixture was dilutedwith EtOAc and washed with water(×3), brine, dried over Na₂SO₄, filteredand concentrated. The crude product was purified on silica gel columnwith hexane/EtOAc, 8:1 to give additional 452 mg of NY-122.

[0806] Using same procedure for NY-110, NY-122 (450 mg, 1.12 mmol) wasconverted to NY-123 (442 mg).

[0807] Using same procedure for 509-HD-213, NY-123 (322 mg, 0.827 mmol)was converted to NY-124 (365 mg).

[0808] Using same procedure for 16, YE-06 (441 mg, 0.598 mmol) wasconverted to crude product of NY-125 (857 mg).

[0809] Using same procedure for 18, NY-125 (857 mg) was converted toNY-126 (231 mg).

[0810] Using same procedure for 509-HD-116, NY-126 (230 mg, 0.233 mmol)was converted to NY-127 (197 mg). NY-127 was used without purificationfor the next step.

[0811] Using same procedure for TM-12, NY-127 (197 mg, 0.233 mmol) wasconverted to NY-128 (53 mg).

[0812] To a solution of NY-₁₂₈ (7.2 mg, 0.0139 mmol) in THF (1 mL),n-BU3P (3.5 μL) was added and stirred at room temperature for 80 min.Additional n-BU₃P (3.5 μL) was added and the mixture was stirred for 80min. Then, water (50 μL) was added to the reaction mixture, and themixture was stirred at room temperature for 3 hrs. The reaction mixturewas dried over Na₂SO₄, filtered and concentrated. The crude product waspurified on silica gel column with CH₂Cl₂/MeOH, 98:2, 95:5, 9:1 to give5.2 mg of NY-129.

[0813] To a solution of NY-129 (5.2 mg, 0.0106 mmol), Et₃N (5 μL, 0.0359mmol), Ac₂O (1.5 μL, 0.0159 mmol) was added at 0° C. and stirred for 40min. The reaction mixture was quenched with sat. NH₄Cl and extractedwith EtOAc. The organic layer was washed with water, sat. NaHCO₃, brine,dried over Na₂SO₄, filtered and concentrated to give 5 mg of NY-130.

[0814] Using same procedure for 509-HD-125, NY-130 (5 mg, 0.00937 mmol)was converted to NY-131 (6.5 mg). NY-131 was used without purificationfor the next step.

[0815] Using same procedure for B2538, NY-131 (6.4 mg, 0.00937 mmol) wasconverted to NF-2553 (1.3 mg).

[0816] Synthetic Procedure for NF-2556

[0817] Using same procedure for NY-121, NY-120 (670 mg, 2 mmol) wasconverted to NY-132 (875 mg).

[0818] Using same procedure for NY-122, NY-132 (873 mg, 2 mmol) wasconverted to NY-133 (716 mg).

[0819] Using same procedure for NY-123, NY-133 (708 mg, 1.7 mmol) wasconverted to NY-134 (694 mg).

[0820] Using same procedure for NY-124, NY-134 (694 mg, 1.7 mmol) wasconverted to NY-135 (799 mg).

[0821] Using same procedure for 16, YE-06 (318 mg, 0.432 mmol) wasconverted to crude product of NY-136 (683 mg).

[0822] Using same procedure for 18, NY-136 (682 mg) was converted toNY-126 (229 mg).

[0823] Using same procedure for 509-HD-116, NY-137 (225 mg, 0.224 mmol)was converted to NY-138 (207 mg). NY-138 was used without purificationfor the next step.

[0824] Using same procedure for TM-12, NY-138 (207 mg, 0.224 mmol) wasconverted to NY-139 (72 mg).

[0825] Using same procedure for NY-129, NY-139 (33 mg, 0.0621 mmol) wasconverted to NY-140 (28.5 mg).

[0826] Using same procedure for NY-130, NY-140 (28 mg, 0.0554 mmol) wasconverted to NY-141 (29.8 mg).

[0827] Using same procedure for 509-HD-125, NY-130 (5 mg, 0.00937 mmol)was converted to NY-131 (18 mg).

[0828] Using same procedure for B2538, NY-143 (18 mg, 0.033 mmol) wasconverted to NF-2556 (12.6 mg).

[0829] Preparation of C13-C Analogs, NF1774, NF2546, NF2550, NF2551,NF2552, NF2554, NF2555, and NF2560

[0830] Synthetic Procedure for NF-1774

[0831] Using same procedure for 9, NY-37 (8.1 g, 33.05 mmol) wasconverted to NY-38 (9.28 g).

[0832] NY-38 (31.76 g, 88.39 mmol) was dissolved in Et₂O (300 mL) andcooled to −78° C., under nitrogen. Then, t-BuLi (1.7M/pentane, 100 mL,170 mmol) was slowly added and the reaction was stirred at −78° C. for20 min. Dry ice was added to the solution, then the solution was allowedto warm to rt and was stirred for 1.5 hrs. The mixture was quenched withsat.NH₄Cl, acidified with 10% citric acid, extracted with EtOAc(×2). Theorganic layers were washed with water, brine and dried over Na₂SO₄,filtered and concentrated to give 28.39 g of NY-39.

[0833] A mixture of NY-39 (25.48 g, 78.54 mmol), dimethyl sulfate (7.8mL, 82.43 mmol), NaHCO₃ (9.9 g, 117.8 mmol) and acetone (200 mL) wasrefluxed for 14 hrs. The insoluble material was filtered and thefiltrate was concentrated. The residue was diluted with EtOAc and washedwith sat.NH₄Cl, water, brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified on silica gel column withhexane/EtOAc, 50:1, 40:1, 30:1, 15:1 to give 7.95 g of NY-40 and 6.97 gof the desilylated product NY-41.

[0834] Using same procedure for NY-07, NY-41 (11.75 g, 52.41 mmol) wasconverted to NY-42 (13.43 g).

[0835] Using same procedure for 10, NY-42 (13.43 g, 50.44 mmol) wasconverted to NY-43 (8.74 g) and the deacetylated product NY-44 (7.86 g).

[0836] A mixture of NY-43 (8.74 g, 23.34 mmol), NY-44 (7.86 g, 23.65mmol), 2N NaOH (230 mL) and MeOH (300 mL) was refluxed for 24 hrs. Themixture was concentrated and the residue was acidified with 2N HCl,extracted with EtOAc. The organic layer was washed with water, brine,dried over Na₂SO₄, filtered and concentrated to give 12.62 g of NY-45.

[0837] A mixture of NY-45 (12.62 g, 41.47 mmol), conc.sulfuric acid (0.8mL) and MeOH (200 mL) was refluxed for 15 hrs. The mixture wasconcentrated and the residue was diluted with EtOAc and washed withwater, brine, dried over MgSO₄, filtered and concentrated. The crudeproduct was purified on silica gel column with hexane/EtOAc, 6:1, 5:1,4:1, 1:1 to give 2 g of NY-46 and 11.1 g of mixture of NY-47 (smallamount) and NY-48.

[0838] Using same procedure for 509-HD-213, a mixture of NY-47 and NY-48(500 mg, 1.57 mmol) was converted to NY-49 (297 mg).

[0839] Using same procedure for 509-HD-209, NY-49 (297 mg, 0.71 mmol)was converted to NY-50 (250 mg).

[0840] To a mixture of 60% NaH in mineral oil (330 mg, 8.25 mmol) andDMF (20 mL), a solution of NY-24 (3.46 g, 5.94 mmol) in DMF (20 mL) wasgradually added at 0° C. and stirred for 30 min. Then, MPMCl (1.2 ml,8.85 mmol) was added and the reaction mixture was allowed to warm toroom temperature and stirred for 12 hrs. The reaction mixture was pouredinto ice-cooled sat. NH₄Cl and extracted with EtOAc. The organic layerwas washed with water (×2), brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified on silica gel column withhexane/EtOAc, 18:1, 15:1 to give 1.9 g of NY-51.

[0841] Using same procedure for NY-26, NY-51 (1.9 g, 2.7 mmol) wasconverted to NY-52 (1.64 g).

[0842] Using same procedure for 554-RB-260, NY-52 (1.64 g, 2.65 mmol)was converted to NY-53 (1.68 g).

[0843] Using same procedure for 16, NY-53 (540 mg, 0.741 mmol) wasconverted to NY-54 (465 mg).

[0844] Using same procedure for 18, NY-54 (458 mg, 0.431 mmol) wasconverted to NY-55 (274 mg).

[0845] Using same procedure for 509-HD-116, NY-55 (273 mg, 0.286 mmol)was converted to NY-56 (166 mg).

[0846] Using same procedure for 509-HD-118, NY-56 (164 mg, 0.267 mmol)was converted to NY-57 (86 mg).

[0847] Using same procedure for 509-HD-188, NY-57 (30 mg, 0.05 mmol) wasconverted to NY-58 (20 mg).

[0848] To a solution of NY-58 (19 mg, 0.04 mmol) in CH₂Cl₂ (5 mL), MS4A(50 mg) PDC (33 mg, 0.088 mmol) were added and the mixture was stirredat room temperature for 12 hrs. The insoluble material was filtered,washed with EtOAc and the filtrate was concentrated. The residue wasdissolved in CH₂Cl₂ (5 mL), then Dess-Martin periodinate (50 mg, 0.118mmol) was added and the mixturte was stirred for 4 days. Sat. NaHCO₃ and10% Na₂S₂O₃ were added and the mixture was extracted with EtOAc. Theorganic layer was washed with water, brine and dried over Na₂SO₄,filtered and concentrated. The crude product was purified on silica gelcolumn with hexane/EtOAc, 5:1, 4:1, 3:1,2:1 to give 6.8 mg of NY-59.

[0849] To a stirred solution of NY-59 (6.8 mg, 0.014 mmol) in THF (0.4mL)-H₂O (0.2 mL) was added trifluoroacetic acid (0.4 mL) at 0° C. Themixture was then allowed to warm to rt. After 2 hrs, the mixture waspoured into a saturated solution of NaHCO₃ and extracted with EtOAc. Theorganic layer was washed with water, brine and dried over anhydrousNa₂SO₄, filtered and concentrated. The residue was dissolved in MeCN(1.2 ml) and then 50% HF (0.2 mL) was added at 0° C. The mixture wasthen allowed to warm to rt. The crude product was purified on silica gelcolumn with hexane/EtOAc, 1:1, 1:2 to give 2.8 mg of NF-1774.

[0850] Synthetic Procedure for NF-2546

[0851] Using same procedure for 509-HD-213, Methyl 2,5-dihydroxybenzoate(8.41 g, 50 mmol) was converted to NY-99 (5.06 g).

[0852] Using same procedure for 509-HD-209, NY-99 (5.4 g, 20.91 mmol)was converted to NY-100 (5.83 g).

[0853] A mixture of NY-100 (5.82 g, 19.25 mmol), 10% Pd on carbon (610mg) and MeOH (100 ml) was hydrogenated at 4 atom for 3 hrs. The catalystwas filtered and the filtrate was concentrated to give 4.16 g of NY-101.It was used without purification for the next step.

[0854] Using same procedure for 611-MS-88, NY-101 (4.15 g, 19.25 mmol)was converted to NY-102 (5.65 g).

[0855] To a mixture of NY-102 (5.64 g, 16.38 mmol), PhB(OH)₂ (4 g, 32.81mmol), K₂CO₃ (3.4 g, 24.6 mmol) and toluene (120 mL) under N₂ bubbling,Pd(Ph₃P)₄ (570 mg, 0.493 mmol) was added and the mixtute was graduallywarmed to 90° C. and stirred for 2 hrs. The reaction mixture wasfiltered and the filtrate was extracted with EtOAc. The organic layerwas washed with sat. NaHCO₃, brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified on silica gel column withhexane/EtOAc, 20:1, 15:1, 12:1 to give 4.14 g of NY-103.

[0856] Using same procedure for NY-85, NY-103 (4.14 g, 15.2 mmol) wasconverted to NY-104 (2.75 g).

[0857] Using same procedure for NY-86, NY-104 (2.7 g, 8.61 mmol) wasconverted to NY-105 (822 mg).

[0858] Using same procedure for NY-87, NY-105 (810 mg, 2.37 mmol) wasconverted to NY-106 (689 mg).

[0859] A mixture of NY-106 (682 mg, 1.986 mmol), CSA (22 mg, 0.095 mmol)and toluene (15 mL) was refluxed for 4 hrs. The reaction mixture wasconcentrated, then the crude product was purified on silica gel columnwith hexane/EtOAc, 10:1, 6:1 to give 471 mg of NY-107.

[0860] Using same procedure for NY-90, NY-107 (470 mg, 1.74 mmol) wasconverted to NY-108 (500 mg). NY-108 was used without purification forthe next step.

[0861] Using same procedure for NY-90, NY-107 (470 mg, 1.74 mmol) wasconverted to NY-108 (500 mg). NY-108 was used without purification forthe next step.

[0862] Using same procedure for NY-91, NY-108 (470 mg, 1.74 mmol) wasconverted to NY-109 (336 mg).

[0863] A mixture of NY-109 (330 mg, 0.837 mmol), KOH (142 mg, 2.53mmol), water (5 mL) and DMSO (15 mL) was heated at 70° C. for 24 hrs.The reaction mixture was diluted with EtOAc and washed with 10% KHSO₄,water, brine, dried over Na₂SO₄, filtered and concentrated to give 334mg of NY-110. NY-110 was used without purification for the next step.

[0864] Using same procedure for 509-HD-213, NY-110 (333 mg, 0.837 mmol)was converted to NY-111 (359 mg).

[0865] Using same procedure for 16, YE-06 (394 mg, 0.535 mmol) wasconverted to crude product of NY-112 (687 mg).

[0866] Using same procedure for 18, NY-112 (682 mg) was converted toNY-113 (215 mg).

[0867] Using same procedure for 509-HD-116, NY-113 (236 mg, 0.241 mmol)was converted to NY-114 (198 mg). NY-114 was used without purificationfor the next step.

[0868] Using same procedure for TM-12, NY-114 (198 mg, 0.241 mmol) wasconverted to NY-115 (31 mg).

[0869] Using same procedure for 509-HD-125, NY-115 (43 mg, 0.0845 mmol)was converted to NY-116 (47 mg).

[0870] Using same procedure for B2538, NY-116 (45 mg, 0.0845 mmol) wasconverted to NF-2546 (29.4 mg).

[0871] Synthetic Procedure for NF2548

[0872] TM-51 (1.0 g, 2.0 mmol) was dissolved in triethylamine (25 mL).MK-123 (0.55 g, 3.1 mmol), PdCl₂(PPh₃)₂ (220 mg, 0.31 mmol), CuI (120mg, 0.63 mmol) were added and the mixture was stirred at 70° C., whileaddition of MK-123 (0.25 g , 1.4 mmol/each time) were repeated atintervals of 1 hr for five times. The mixture was cooled to rt andconcentrated. The concentrate was triturated with diethyl ether andhexane, and the insoluble solid was filtered off. The filtrate wasconcentrated and the residue was purified by chromatography on silicagel using 5-10% EtOAc/hexane to give 0.67 g (1.2 mmol, 56%) of YE-32.

[0873] Using similar procedure for the intermediate 18 of NF2561, theiodide (430 mg, 0.58 mmol) was converted to YE-33 (430 mg, 0.40 mmol,68% 3 steps).

[0874] Using similar procedure for TM-12, YE-33 (430 mg, 0.40 mmol) wasconverted to YE-34 (113 mg, 0.19 mmol, 47%, 3 steps).

[0875] Using similar procedure for TM-13, YE-34 (40 mg, 0.066 mmol) wasconverted to enone. The following deprotection of the enone by DDQresulted in deprotection of both MPM and MOM group and provided 16 mg ofYE-35 (0.036 mmol, 55% 2 steps).

[0876] Using similar procedure for YE-19, YE-35 (12 mg, 0.027 mmol) wasconverted to NF-2548 (10.5 mg, 0.026 mmol, 96%).

[0877] Synthetic Procedure for NF2549

[0878] YE-35 (4 mg, 0.0091 mmol) and triethylamine (0.01 mL, 0.071 mmol)were dissolved in THF (0.6 mL). Methyl isocyanate (total 0.070 mL, 1.2mmol) was added in four portions in 2 h intervals at rt. After 4 days,excess isocyanate was quenched with ethyleneglycol (0.15 ml). Themixture was poured into water and extracted with EtOAc. The extract waswashed with 5% KHSO₄ aq., a saturated NaHCO₃ sol., brine, dried overanhydrous Na₂SO₄, filtered and concentrated. The crude product waspurified by chromatography on silica gel using 15-30% EtOAc/hexane togive 4 mg (0.0080 mmol, 88%) of the N-methylcarbamate.

[0879] Using similar procedure for ER803064, the N-methylcarbamate (4mg, 0.0080 mmol) was converted to NF2549 (3 mg, 0.0028 mmol, 35%).

[0880] Synthetic Procedure for NF2554, NF2555

[0881] Using similar procedure for YE-32, the TM-49 (2.2 g, 8.0 mmol)was converted to YE-36 (2.1 g, 5.8 mmol, 72%).

[0882] To a stirred solution of YE-36 (2.1 g, 5.7 mmol) in EtOH (60 mL)and THF (15 mL) was added Raney Ni W-2 (water susupension, 4.5 g) undernitrogen atmosphere. The suspension was stirred under hydrogenatmosphere overnight. Then the catalyst was removed by filtration, andthe filtrate was concentrated. The concentrate was purified bychromatography on silica gel using 15-33% EtOAc/hexane to give 1.6 g(4.3 mmol, 75%) of YE-37.

[0883] Using similar procedure for TM-51, YE-37 (1.18 g, 3.2 mmol) wasconverted to YE-38 (0.20 g, 0.40 mmol, 13%, 3steps).

[0884] Using similar procedure for TM-39, YE-38 (340 mg, 0.68 mmol) wasconverted to YE-39 (350 mg, 0.64 mmol, 88% 2 steps).

[0885] Using similar procedure for the intermediate 18 of NF2561, theiodide (230 mg, 0.30 mmol) was converted to YE-40 (115 mg, 0.11 mmol,34% 3 steps).

[0886] Using similar procedure for TM-12, YE-40 (115 mg, 0.11 mmol) wasconverted to YE-41 (36 mg, 0.059 mmol, 55% 3 steps).

[0887] Using similar procedure for TM-13 followed by usual deprotectionof MPM group by DDQ oxidation, YE-41 (36 mg, 0.059 mmol) was convertedto YE-42 (13 mg, 0.027 mmol, 45% 2 steps).

[0888] Using similar procedure for ER803064, YE-42 (5 mg, 0.010 mmol)was converted to NF2554 (4.1 mg, 0.010 mmol, 99%).

[0889] Using similar procedure for NF-2549, YE-42 (7 mg, 0.014 mmol) wasconverted to NF-2555 (1.9 mg, 0.0041 mmol, 29% 2 steps).

[0890] Synthetic Procedure for NF2550

[0891] Using similar procedure for 509-HD-207, starting material (4.91g, 25 mmol) was converted to TM-34 (5.17 g, 18 mmol, 72%).

[0892] To a stirred suspension of NaH (60% oil dispersion, 593 mg, 14.8mmol) in dry DMF (5 mL) was added a solution of TM-34 (3.395 g, 11.9mmol) in dry DMF (35 mL) at 0° C. under nitrogen atmosphere. After 30min, methyl iodide (3.7 mL, 59 mmol) was added. the mixture was allowedto warm to rt and stirred for 30 min after which a saturated solution ofNH₄Cl was added. The mixture was extracted with EtOAc and the organicextract was washed with water, brine, dried with anhydrous Na₂SO₄,filtered and concentrated.

[0893] The crude product (4.33 g) was dissolved in EtOH (60 mL). Then,10% Pd on carbon (630 mg) was added. Reaction was stirred underhydrogen. After 3 hrs, reaction was stopped, catalyst was filteredthrough celite and mixture was concentrated under reduced pressure togive the phenol (2.94 g) as crude product.

[0894] To a solution of the phenol (2.94 g) in dry DMF (60 mL) wereadded anhydrous K₂CO₃ (3.28 g, 24 mmol) and 2-bromo-1,1-diethoxyethane(2.86 mL, 18 mmol) and the mixture was stirred for 5 hrs at 140° C.After cooling to rt, the mixture was poured into water and extractedwith EtOAc. The organic extract was washed with a saturated solution ofNH₄Cl, water, brine, dried with anhydrous Na₂SO₄, filtered andconcentrated. The crude product was purified by chromatography on silicagel using 15% EtOAc/hexane to give 3.369 g (10 mmol, 87% 3 steps) ofTM-35.

[0895] To a solution of TM-35 (3.02 g, 9.3 mmol) in toluene (120 mL) wasadded PPA (5.9 mL) and the mixture was stirred for 2 hrs at 100° C.under nitrogen atmosphere. After cooling to rt, the mixture was pouredinto ice/water and extracted with EtOAc. The organic extract was washedwith water, a saturated solution of NaHCO₃, brine, dried with anhydrousNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel using 12.5% EtOAc/hexane to give 395 mg(1.7 mmol, 18%) of TM-36.

[0896] To a stirred solution of TM-36 (541 mg, 2.3 mmol) in dry CH₂Cl₂(20 mL) was added BBr₃ (0.44 mL, 4.6 mmol) at −78° C. under nitrogenatmosphere. After 1 hr, the mixture was poured into water and extractedwith EtOAc. The organic extract was washed with a saturated solution ofNaHCO₃, water, brine, dried with anhydrous Na₂SO₄, filtered andconcentrated. The crude product was purified by chromatography on silicagel using 5% EtOAc/hexane to give 357 mg (1.6 mmol, 70%) of the phenol.

[0897] To a stirred solution of the phenol (170 mg, 0.77 mmol) in dryTHF (8 mL) were added DBU (0.23 mL, 1.5 mmol) and chloromethyl methylether (0.12 mL, 1.5 mmol) at rt under nitrogen atmosphere. After 3 hr,the mixture was poured into a saturated solution of NH₄Cl and extractedwith EtOAc. The organic extract was washed with 10% KHSO₄ aq., water, asaturated solution of NaHCO₃, brine, dried with anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 15% EtOAc/hexane to give 193 mg (0.73mmol, 95%) of TM-37.

[0898] Using similar procedure for the intermediate 10 of NF2561, TM-37(192 mg, 0.73 mmol) was converted to TM-38 (192 mg, 0.52 mmol, 71% 2steps).

[0899] Using similar procedure for the intermediate 14 of NF2561, TM-38(192 mg, 0.52 mmol) was converted to TM-39 (190 mg, 0.43 mmol, 83% 2steps).

[0900] Using similar procedure for the intermediate 18 of NF2561, theiodide (223 mg, 0.30 mmol) was converted to TM-40 (232 mg, 0.25 mmol,81% 3 steps).

[0901] Using similar procedure for TM-12, TM-40 (232 mg, 0.25 mmol) wasconverted to TM-41 (81 mg, 0.17 mmol, 70% 3 steps).

[0902] Using similar procedure for ER803064, TM-41 (41 mg, 0.087 mmol)was converted to NF2550 (20 mg, 0.052 mmol, 60% 2 steps).

[0903] Synthetic Procedure for NF2560

[0904] To a stirred solution of 2,2,6,6-tetramethylpiperidine (0.05 mL,0.3 mmol) in dry THF (1.5 mL) was added 1.6M n-BuLi in hexane (0.15 mL,0.24 mmol) at −20° C. under nitrogen atmosphere. After 30 min, themixture was cooled to −78° C. and a solution of TM-41 (38 mg, 0.08 mmol)in dry THF (4 mL) was added. After 1 hr, dry DMF (0.06 mL) was added andthe mixture was allowed to warm to rt. It was quenched with a saturatedsolution of NH₄Cl and extracted with EtOAc. The organic extract waswashed with a saturated solution of NH₄Cl, water, brine, dried withanhydrous Na₂SO₄, filtered and concentrated. The crude product waspurified by chromatography on silica gel using 40% EtOAc/hexane to give18 mg (0.036 mmol, 45%) of TM-42.

[0905] To a stirred solution of TM-42 (54 mg, 0.11 mmol) in CH₂Cl₂ (5mL) were added triethylamine (0.3 mL, 2.2 mmol), small amount ofN,N-dimethylaminopyridine, acetic anhydride (0.1 mL, 1.1 mmol) at 0° C.The mixture was allowed to warn to rt and stirred overnight. It wasquenched with a saturated solution of NH₄Cl and extracted with EtOAc.The organic extract was washed with a saturated solution of NH₄Cl,water, brine, dried with anhydrous Na₂SO₄, filtered and concentrated.

[0906] The crude acetate (61 mg) was dissolved in MeOH (5 mL) and NaBH₄(10 mg, 0.26 mmol) was added at 0° C. After 10 min, the mixture waspoured into a saturated solution of NH₄Cl and extracted with EtOAc. Theorganic extract was washed with a saturated solution of NH₄Cl, water,brine, dried with anhydrous Na₂SO₄, filtered and concentrated. The crudeproduct was purified by chromatography on silica gel using 50%EtOAc/hexane to give 40 mg (0.073 mmol, 68% 2 steps) of TM-43.

[0907] To a stirred solution of TM-43 (40 mg, 0.073 mmol) in dry THF (8mL) were added triphenylphosphine (58 mg, 0.22 mmol), diphenylphosphorylazide (0.047 mL, 0.22 mmol), 40% DEAD in toluene (0.1 mL, 0.22 mmol) atrt. After 20 hrs, the mixture was concentrated under reduced pressureand the residue was purified by chromatography on silica gel using 25%EtOAc/hexane to give 26 mg (0.046 mmol, 62%) of TM-44.

[0908] To a stirred solution of TM-44 (26 mg, 0.046 mmol) in dry THF(2.5 mL) was added tributylphosphine (0.035 mL, 0.14 mmol) at rt undernitrogen atmosphere. After 30 min, H₂O (0.5 mL) was added. Afteradditional 3 hrs, the mixture was diluted with EtOAc and dried withanhydrous Na₂SO₄, filtered and concentrated.

[0909] The crude amine (56 mg) was dissolved in CH₂Cl₂ (3 mL). Thentriethylamine (0.036 mL, 0.26 mmol) and methanesulfonyl chloride (0.010mL, 0.13 mmol) were added at 0° C. After 30 min, the mixture was pouredinto a saturated solution of NaHCO₃ and extracted with EtOAc. Theorganic extract was washed with a saturated solution of NaHCO₃, water,brine, dried with anhydrous Na₂SO₄, filtered and concentrated.

[0910] The crude methanesulfonamide (64 mg) was dissolved in EtOH (3 mL)and 1N NaOH aq. (0.5 mL, 0.5 mmol) was added at rt. After 24 hrs, themixture was quenched with 1N HCl (0.5 mL) and extracted with EtOAc. Theorganic extract was washed with water, brine, dried with anhydrousNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel using 60% EtOAc/hexane to give 15 mg (0.026mmol, 57% 3 steps) of TM-45.

[0911] Using similar procedure for ER803064, TM-45 (15 mg, 0.026 mmol)was converted to NF2560 (4.4 mg, 0.0089 mmol, 35% 2 steps).

[0912] Synthetic Procedure for NF2545

[0913] To a stirred suspension of 3-methoxybenzyl alcohol (10.78 g, 78mmol) in hexane (100 mL) was added 1.6M n-BuLi in hexane (100 mL, 160mmol) at −30° C. under nitrogen atmosphere. After 1.5 hrs, dry CO₂ wasbubbled for 30 min and then the mixture was poured into water andextracted with diethyl ether. The aqueous layer was acidified byaddition of 5N HCl at 0° C. and stirred overnight at rt. The precipitatewas collected by filtration and washed with water, dried at 70° C. for24 hrs to give 4.79 g (29 mmol, 37%) of TM-46.

[0914] TM-46 (4.79 g, 29 mmol) was dissolved in CH₂Cl₂ (150 mL). Thenbromine (1.88 mL, 37 mmol) was added at 0° C. and the mixture wasallowed to warm to rt. After 20 hrs, the mixture was poured into asaturated solution of Na₂S₂O₃ and extracted with EtOAc. The organicextract was washed with a saturated solution of NaHCO₃, water, brine,dried with anhydrous Na₂SO₄, filtered and concentrated to give 6.59 g(27 mmol, 93%) of TM-47.

[0915] Using similar procedure for TM-37, TM-47 (5.37 g, 22 mmol) wasconverted to TM-49 (5.98 g, 22 mmol, 99% 2 steps).

[0916] To a stirred solution of TM-49 (3.85 g, 14 mmol) in DMSO (40 mL)was added a solution of KOH (851 mg, 15 mmol) in H₂O (15 mL). After 30min, H₂O was removed by evaporation and methyl iodide (2.63 mL, 42 mmol)was added and the mixture was stirred for 30 min. The mixture wasdiluted with EtOAc and washed with water, a saturated solution ofNaHCO₃, water, brine, dried with anhydrous Na₂SO₄, filtered andconcentrated.

[0917] The crude product (3.79 g) was dissolved in dry DMF (100 mL).Then diphenyl disulfide (6.16 g, 28 mmol), pyridine (4.55 mL, 56 mmol),tributylphosphine (7.03 mL, 28 mmol) were added at rt under nitrogenatmosphere and the mixture was stirred overnight. It was quenched with asaturated solution of NaHCO₃ and extracted with EtOAc. The organic

[0918] extract was washed with 10% KHSO₄ aq., water, brine, dried withanhydrous Na₂SO₄, filtered and concentrated. The crude product waspurified by chromatography on silica gel using 15% EtOAc/hexane to give1.79 g (4.5 mmol, 32% 2 steps) of TM-50.

[0919] Using similar procedure for TM-39, TM-50 (1.79 g, 4.5 mmol) wasconverted to TM-51 (1.88 g, 3.9 mmol, 87% 2 steps).

[0920] TM-51 (1.86 g, 3.8 mmol) was dissolved in DMF (30 mL). Thentriethylamine (10 mL), 3-butyn-1-ol (1.16 mL, 15 mmol), PdCl₂(PPh₃)₂(404 mg, 0.58 mmol), CuI (219 mg, 1.2 mmol) were added and the mixturewas stirred overnight at 70° C. The mixture was diluted with 50%EtOAc/hexane and washed with water, brine, dried with anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 30% EtOAc/hexane to give 1.76 g (3.7mmol, 97%) of TM-52.

[0921] Using similar procedure for TM-45, TM-52 (754 mg, 1.6 mmol) wasconverted to TM-53 (586 mg, 1.1 mmol, 67% 3 steps).

[0922] Using similar procedure for TM-39, the iodide (350 mg, 0.48 mmol)was converted to TM-54 (224 mg, 0.25 mmol, 45% 3 steps).

[0923] Using similar procedure for NF2550, TM-54 (224 mg, 0.21 mmol) wasconverted to NF2545 (2.8 mg, 0.0057 mmol, 2.7% 5 steps).

[0924] Synthetic Procedure for NF2551 and NF2552

[0925] Using similar procedure for TM-52 followed by hydrogenation,TM-49 (2.95 g, 11 mmol) was converted to TM-55 (2.41 g, 9.1 mmol, 84% 2steps).

[0926] To a stirred solution of TM-55 (2.41 g, 9.1 mmol) in cyclohexane(30 mL)-CH₂Cl₂ (15 mL) were added CSA (105 mg, 0.45 mmol) and4-methoxybenzyl trichloroacetimidate (7.68 g, 27 mmol) at rt undernitrogen atmosphere. After 14 hrs, triethylamine (0.1 mL) was added.Then precipitate was removed by filtration and washed with hexane. Thefiltrate was concentrated under reduced pressure. The residue waspurified by chromatography on silica gel using 30% EtOAc/hexane to give3.26 g (8.4 mmol, 93%) of TM-56.

[0927] Using similar procedure for TM-50, TM-56 (3.26 g, 8.4 mmol) wasconverted to TM-57 (1.78 g, 3.5 mmol, 41% 3 steps).

[0928] Using similar procedure for TM-39, TM-57 (366 mg, 0.72 mmol) wasconverted to TM-58 (381 mg, 0.64 mmol, 89% 2 steps).

[0929] Using similar procedure for TM-39, the iodide (223 mg, 0.30 mmol)was converted to TM-59 (132 mg, 0.12 mmol, 40% 3 steps).

[0930] Using similar procedure for TM-12, TM-59 (132 mg, 0.12 mmol) wasconverted to TM-60 (32 mg, 0.051 mmol, 43% 3 steps).

[0931] Using similar procedure for TM-13 followed by usual deprotectionof MPM group by DDQ oxidation, TM-60 (32 mg, 0.051 mmol) was convertedto TM-61 (15 mg, 0.030 mmol, 58% 2 steps).

[0932] Using similar procedure for ER803064, TM-61 (8 mg, 0.016 mmol)was converted to NF2551 (4 mg, 0.010 mmol, 60%).

[0933] To a stirred solution of TM-61 (7 mg, 0.014 mmol) in CH₂Cl₂ (1mL) were added triethylamine (0.02 mL) and methyl isocyanate (0.1 mL) atrt. After 15 hrs, the mixture was diluted with EtOAc and washed with 3%NH₄OH aq., 5% KHSO₄ aq., water, brine, dried with anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 55% EtOAc/hexane to give 7 mg (0.013mmol, 90%) of the N-methylcarbamate.

[0934] Using similar procedure for ER803064, the N-methylcarbamate (7mg, 0.013 mmol) was converted to NF2552 (3 mg, 0.0063 mmol, 50%).

[0935] Preparation of the Intermediate for C14-O-Linked Including C2-C4Analogs:

[0936] Known compound 1 (0.22 g, 1.2 mmol) and K₂CO₃ (0.25 g, 1.8 mmol)was dissolved in 12 mL of DMF. After the addition of MPM-Cl (0.17 mL,1.2 mmol) the mixture was heated at 35° C. for 12 hours. Crude mixturewas concentrated and filtered. The resulted material was purified bysilica gel chromatography. CH₂Cl₂ was used to recover 445-ycs-252 (0.16g, 0.53 mmol) from silica gel plug in 44% yield.

[0937] 60% of NaH in mineral oil (16 mg, 0.40 mmol) was added to445-ycs-252 (36 mg, 0.12 mmol) in 2 mL of DMF. After the addition ofMOMCl (0.17 μL, 0.22 mmol), the mixture was stirred at rt for 1 hour.DMF was evaporated under high vacuum, and the residue was dissolved inCH₂Cl₂, washed with H₂O. The crude material was purified by silica gelchromatography. 30% EtOAc in hexanes was used to elute 445-ycs-254 (40mg, 0.12 mmol) from silica gel plug in 97% yield.

[0938] 2.5 M n-BuLi in hexanes (13 mL, 33 mmol) was introduced drop wiseto the stirring solution of diisopropylamine (4.6 mL, 33 mmol) in 30 mLof THF at −5° C. The solution was stirred at 0° C. for 30 minutes beforeit was cooled to −78° C. 445-ycs-254 (6.3 g, 18 mmol) in 25 mL of THFwas added to the cold LDA slowly so that the internal temperature waskept below −78° C. The mixture was stirred at −78° C. for 45 minutes,and then (PhSe)₂ (5.2 g, 17 mmol) in 25 mL of THF was added slowly sothat the internal temperature was kept below −60° C. Reaction mixturewas stirred for 40 minutes before it was quenched with aq. NH₄Cl at −78°C. EtOAc was added to the mixture at rt. After separation, organic layerwas dried (Na₂SO₄) and concentrated. 5% EtOAc in toluene was used toelute 445-ycs-268 (6.9 g, 14 mmol) from silica gel column in 75% yield.

[0939] 2.5 N NaOH (18 mL, 46 mmol) was added to 445-ycs-268 (7.7 g, 15mmol) in 18 mL of EtOH. The mixture was refluxed for 12 hours before itwas acidified and extracted with EtOAc. Organic phase was dried (Na₂SO₄)and concentrated to afford white crystalline 445-ycs-272 (7.2 g, 15mmol) in 96% yield.

[0940] DEAD (3.5 mL, 22 mmol) was added to a solution of 445-ycs-272(7.2 g, 15 mmol), Ph₃P (5.8 g, 22 mmol) and 2-TMS-ethanol (2.6 mL, 18mmol) in 200 mL of toluene at 0° C. The mixture was stirred at RT for 1hour before it was quenched with aq. NaHCO₃. Organic phase was dried(Na₂SO₄), concentrated and purified by silica gel chromatography toafford 445-ycs-273 (8.2 g, 14 mmol) in 95% yield.

[0941] A procedure similar to the preparation of 554-RB-260 was used.

[0942] LiHMDS in THF (2.8 mL, 2.8 mmol) was introduced drop wise into acold (−78° C.) solution of 445-ycs-274 (1.4 g, 1.9 mmol) and 445-ycs-273(1.7 g, 2.8 mmol) in 10 mL of 10:1 THF-HMPA. The internal temperaturewas kept below −70° C. during the addition. The mixture was stirred at−78° C. for half hour before it was quenched with aq. NH₄Cl and dilutedwith EtOAc. Organic phase was dried (Na₂SO₄), concentrated and purifiedby silica gel chromatography to afford 445-ycs-278 (1.9 g, 1.6 mmol) in82% yield.

[0943] m-CPBA (1.0 g, 4.2 mmol) was added in three portions into a cold(0° C.) solution of 445-ycs-278 (2.5 g, 2.1 mmol) in 30 mL of CH₂Cl₂.The mixture was stirred at 0° C. for 1 hour before the addition of Et₃N(1.8 mL, 13 mmol). After the reaction mixture was stirred at rt for 1hour it was quenched with aq. Na₂S₂O₃ and diluted with aq. NaHCO₃.Organic phase was dried (Na₂SO₄), concentrated and purified by silicagel chromatography to afford 445-ycs-281 (1.6 g, 1.6 mmol) in 76% yield.

[0944] In a solution of TBAF buffered with 0.33 mole equivalent ofimidazole.HCl (1.3 mL, 1.3 mmol) was introduced to the solution of445-ycs-281 (0.17 g, 0.17 mmol) in 2 mL of THF. The mixture was stirredat 50° C. for 12 hours before it was diluted with Et₂O and washed withaq. NH₄Cl. Organic phase was dried (Na₂SO₄) and concentrated to furnishcrude 445-ycs-295, which dissolved in 20 mL of CH₂Cl₂ was added dropwise to the refluxing mixture of 2-chloro-1-methylpyridinium iodide(0.12 g, 0.48 mmol) and n-Bu₃N (0.11 mL, 0.48 mmol) in 12 mL of CH₂Cl₂.The mixture was refluxed for 2 hours before it was diluted with Et₂O andwashed with 0.05 N HCl, H₂O and NaHCO₃. Organic phase was dried(Na₂SO₄), concentrated and purified by silica gel chromatography toafford 445-ycs-299 (90 mg, 0.13 mmol) in 81% yield for 2 steps.

[0945] p-TsOH.H₂O (11 mg, 0.060 mmol) was added in two portions to asolution of 445-ycs-299 (40 mg, 0.060 mmol) in 4.5 mL of 2:1 MeOH-THF atrt, The solution was stirred at 40° C. for 3 days before it wasconcentrated and purified by silica gel chromatography to afford560-ycs-30 (15 mg, 0.032 mmol) in 53% yield.

[0946] Catalytic amount of p-TsOH.H₂O (1 crystal) was added to thesolution of 560-ycs-30 (13 mg, 0.028 mmol) and excess amount of2-methoxypropane (2 drops) in 1 ML of CH₂Cl₂. The mixture was stirred atRT for 2 hours before the addition of NaHCO₃ and filtration. Thesolution was concentrated and purified by silica gel chromatography toafford 560-ycs-36 (11 mg, 0.022 mmol) in 79% yield.

[0947] Preparation of ER804104: 560-ycs-36 was used as an advancedintermediate for following analogs synthesis as examples.

[0948] 1 N NaOH (2.0 mL, 2.0 mmol) was introduced into a solution of445-ycs-299 (43 mg, 0.064 mmol) in 3 mL of 2:1 EtOH-THF at rt. Thesolution was stirred at 40° C. for 12 hours before it was diluted withEt₂O and brine. Organic phase was dried (Na₂SO₄) and purified by silicagel chromatography to afford 445-ycs-311 (33 mg, 0.058 mmol) in 91%yield.

[0949] PCC (38 mg, 0.17 mmol) was added in three portions to a mixtureof 445-ycs-311 (33 mg, 0.058 mmol), 4A molecular sieves (40 mg) andcelite (40 mg) in 2 mL of CH₂Cl₂ at rt. The mixture was stirred at rtfor 1 hour before it was diluted with Et₂O and filtered. The filtratewas concentrated and passed through a short plug of silica gel (1:1EtOAc-Hexanes) to afford 560-ycs-9 (28 mg, 0.049 mmol) in 86% yield.

[0950] 560-ycs-9 was deprotected as described for the synthesis ofER803064 to give ER804104.

[0951] ER-804168 The synthesis of ER-804168 was the same as thesynthesis of ER-803064

[0952] Synthesis of ER-805125:

[0953] The synthesis of 507-XYL-147 from 507-XYL-111 was followed thesame procedures as in the synthesis of ER-803064.

[0954] To 507-XYL-147 (1.43 g, 1.29 mmol) in a co-solvent of THF/water(5:1 v/v, 60 mL) at room temperature was added OXONE and the mixture wasstirred at room temperature for six hours. The mixture was diluted withethyl acetate and washed three times with water. The organic layer wasdried (sodium sulfate), concentrated and purified by silica gelchromatography to obtain 1.05 g (86%) of the desired product,507-XYL-148 which was confirmed by NMR and MS (M+Na=972).

[0955] The synthesis of 507-XYL-154 from 507-XYL-148 was followed thesame procedures as in the synthesis of ER-803064.

[0956] To 507-XYL-154 (340 mg, 0.57 mmol) in a co-solvent of THF/water(4:1 v/v, 12.5 mL at 0° C. were added N-methyl morpholino-N-oxide (82.5mg, 0.68 mmol) and then osmium tetraoxide in toluene (0.1 M, 0.6 mL,0.06 mmol) in portions. The mixture was stirred at 0° C. for 19 hours.The reaction was quenched with saturated sodium thiosulfate solution anddiluted with ethyl acetate. The mixture was washed with saturated sodiumthiosulfate solution and water. The aqueous layers were back extractedtwice with ethyl acetate and three times with methylene chloride. Thecombined organic layers were dried (sodium sulfate), concentrated andpurified by preparative TLC eluting with 10% methanol in methylenechloride to yield 158 mg (44%) of desired product, 507-XYL-165 which wasconfirmed by NMR and MS (M+Na=649).

[0957] To 507-XYL-165 (120 mg, 0.197 mmol) were added methylene chloride(5 mL), 2,2-dimethoxypropane (0.2 mL, 1.63 mmol) and pyridinium tosylate(12 mg, 0.048 mmol). The mixture was stirred at room temperature for twohours. The mixture was diluted with methylene chloride and washed twicewith saturated sodium bicarbonate solution. The organic layer wasconcentrated and purified by prep TLC to obtain 131 mg of the desiredproducts, 507-XYL-168 which were confirmed by NMR and MS (M+Na=689).

[0958] The remaining synthesis of ER-805125 from 507-XYL-168 wasfollowed the same procedures as in the synthesis of ER-803064.

[0959] Preparation of ER-805216 and ER-805217:

[0960] The starting material 507-XYL-165 (42 mg, 0.066 mmol) wasazeotroped with toluene and dried under high vacuum for one hour. Thestarting material was then dissolved in dry methylene chloride (5 mL)and cooled to 0° C. To this were added collidine (20.3 μL, 0.15 mmol)and then added methanesulfonyl anhydride solution in methylene chloride(0.1 M, 0.69 mL, 0.069 mmol) drop wise over five minutes. The mixturewas stirred at 0° C. for 1.5 hour and at 4° C. over night. The mixturewas poured into a saturated sodium bicarbonate solution and extractedthree times with methylene chloride and once with ethyl acetate. Thecombined organic layers were dried (sodium sulfate), concentrated andpurified by preparative TLC eluting with 5% methanol in methylenechloride to provide 36 mg (77%) of the desired product, 507-XYL-178which was confirmed by NMR and MS (M+Na=727).

[0961] To 507-XYL-178 (36 mg, 0.05 mmol) were added 2.0M ammonia inmethanol (16 mL) and concentrated aqueous ammonium hydroxide solution(3.2 mL) and the mixture was stirred at room temperature for 12 hours.The mixture was concentrated in vacuum, azeotroped with ethyl acetate,methanol and toluene, and dried under reduced pressure. The crudeproduct, 507-XYL-192 was directly used for the next reaction.

[0962] To 507-XYL-192 in methylene chloride (10 mL) at 0° C. were addedtriethylamine (0.2 mL, 1.51 mmol) and acetic anhydride (0.1 mL, 1.06mmol). The mixture was stirred at room temperature over night. Thereaction mixture was cooled to 0° C. and quenched with saturated sodiumbicarbonate solution. The mixture was poured into excessive sodiumbicarbonate solution and extracted three times with methylene chlorideand three times with ethyl acetate. The combined organic layers weredried (sodium sulfate), concentrated to obtain the crude desiredproduct, 507-XYL-194 which was confirmed by MS (M+Na=732). The crudeproduct was directly used for the next reaction without furtherpurification.

[0963] The conversion of 507-XYL-194 to 507-XYL-198 was the same as inthe synthesis of ER-803064. Preparative TLC purified the crude productwith 5% ethanol in ethyl acetate to give 13.6 mg (57% over three steps)of the desired product, 507-XYL-198 which was confirmed by NMR and MS(M+Na=586).

[0964] The oxidation of 507-XYL-198 using pyrridinium chlorochromate wasfollowed the same procedure as in the synthesis of ER-803064.Preparative TLC purified the crude material with 8% ethanol in ethylacetate to afford 507-XYL-204a and 507-XYL-204b, which were confirmed byNMR.

[0965] ER-805216 The conversion of 507-XYL-204a to ER-805216 was thesame as in the synthesis of ER-803064. Purification of crude material bypreparative TLC with 25% ethanol in ethyl acetate afforded ER-805216,which was confirmed by NMR and MS (M+Na=498).

[0966] The conversion of 507-XYL-204b to ER-805217 was the same as inthe synthesis of ER-803064. Purification of crude material bypreparative TLC eluting with 25% ethanol in ethyl acetate affordedER-805217, which was confirmed by NMR and MS (M+Na=500).

[0967] Preparation of ER804401:

[0968] Procedure for the preparation of 560-ycs-86 was similar to thatof 445-ycs-273.

[0969] Procedure for the preparation of 560-ycs-88 was similar to thatof 445-ycs-254.

[0970] Procedure for the preparation of ER804401 from 560-ycs-88 wassimilar to the synthesis of ER804104.

[0971] ER804504 was prepared in the similar way as ER804401 from560-ycs-36 and n-butanol.

[0972] Preparation of ER804555 and ER804567:

[0973] Procedure for the preparation of ER804555 from 560-ycs-36 andN-acetyl ethanolamine was similar to the synthesis of ER804401.

[0974] ER804567 was prepared in the similar way as ER804401 from560-ycs-36 and 3,4-dichlorobenzyl alcohol.

[0975] Preparation of ER804606:

[0976] ER804606 was prepared in the similar way as ER804401 from560-ycs-36 and 4-(2-hydroxyethyl)-morpholine except that oxidation ofalcohol 560-ycs-128 into enone 560-ycs-133 was carried out under Swernconditions as below instead of using PCC.

[0977] (COCl)₂ (2.6 μL, 0.030 mmol) was introduced into a solution ofDMSO (4.2 μL, 0.060 mmol) in 0.2 mL of CH₂Cl₂ at −78° C. The mixture wasstirred at −78° C. for 15 minutes before the addition of 560-ycs-128(6.0 mg, 0.010 mmol) in 0.8 mL of CH₂Cl₂. After the solution was stirredfor 1 hour at −78° C. Et₃N (12 μL, 0.090 mmol) was added. The reactionmixture was allowed to warm up to −20° C. for 15 minutes and quenchedwith Sat. NH₄Cl. Organic phase was washed with NaHCO₃ and concentratedto furnish 560-ycs-133 (6.0 mg, 0.010 mmol) in quantitative yield.

[0978] Preparation of ER804630:

[0979] ER804630 was prepared in the similar way as ER804401 from560-ycs-36 and N-acetyl, N-methyl ethanolamine.

[0980] Preparation of ER804778:

[0981] ER804778 was prepared in the similar way as ER804401 from560-ycs-36 and 3-methylthio-1-propanol except that 560-ycs-199 wasoxidized into 560-ycs-200 by m-CPBA.

[0982] MCPBA (12 mg, 0.048 mmol) was added in three portions to thesolution of 560-ycs-199 (5.2 mg, 0.0081 mmol) in 2 mL of CH₂Cl₂ at 0° C.The solution was stirred at 0° C. for 10 minutes before it was washedwith Na₂S₂O₃ and concentrated to afford 560-ycs-200 (5.0 mg, 0.0074mmol) in 92% yield.

[0983] Aq. HCl (3.0 ML, 3.0 mmol) was introduced to a solution of560-ycs-9 (31 mg, 0.055 mmol) in 3 mL of CH₃CN at rt. The solution wasstirred at rt for 12 hours before it was diluted with EtOAc and H₂O.Organic phase was washed with NaHCO₃, dried (Na₂SO₄), concentrated andpurified by silica gel chromatography to afford ER804104 (12 mg, 0.025mmol) in 45% yield and ER804131 (5.9 mg, 0.016 mmol) in 30% yield.

[0984] Alternative preparation of commonly used intermediate for C14modification:

[0985] Procedure for the preparation of 560-ycs-218 was similar to thatof 445-ycs-273.

[0986] MOM-Cl (2.8 mL, 37 mmol) was added to a solution of 560-ycs-218(2.1 g, 7.4 mmol) and DBU (7.1 mL, 48 mmol) in 10 mL of CH₂Cl₂ at 0° C.The mixture was stirred at rt for 15 minutes before the organic phasewas washed with NaHCO₃, concentrated and purified by silica gelchromatography to furnish 560-ycs-243 (1.8 g, 5.5 mmol) from silica gelplug in 74% yield.

[0987] Procedure for the preparation of 560-ycs-248 was similar to thatof 445-ycs-268.

[0988] Procedure for the preparation of 560-ycs-250 was similar to thatof 445-ycs-272.

[0989] Procedure for the preparation of 560-ycs-251 was similar to thatof 445-ycs-273.

[0990] Procedure for the preparation of 560-ycs-256 was similar to thatof 445-ycs-278.

[0991] Procedure for the preparation of 560-ycs-258 was similar to thatof 445-ycs-281.

[0992] Procedure for the preparation of 560-ycs-267 was similar to thatof 445-ycs-295.

[0993] Procedure for the preparation of 560-ycs-269 was similar to thatof 445-ycs-299.

[0994] TBAF in THF (7.0 mL, 7.0 mmol) was introduced to the solution of560-ycs-269 (0.87 g, 1.3 mmol) in 7 mL of THF. The mixture was stirredat rt for 4 hours before it was diluted with Et₂O and washed with brine.Organic phase was dried (Na₂SO₄), concentrated and purified by silicagel chromatography to afford 560-ycs-270 (0.43 g, 0.78 mmol) in 58%yield over 3 steps.

[0995] Preparation of ER805190:

[0996] Transformations from 560-ycs-279 to 560-ycs-286 and from560-ycs-297 to 560-ycs-300 were similar as the procedure for thepreparation of ER804606.

[0997] Transformations from 560-ycs-286 to 560-ycs-297 and from560-ycs-300 to ER805190 were similar as the procedure for thepreparation of ER805135.

[0998] Preparation of C14-C2 Linked Series:

[0999] To a solution of starting diphenol (10.7 g) in acetone (100 mL),BrCH₂CH₂Cl (15 mL) and 20 g of K₂CO₃ were added. The mixture was heatedat 80° C. for 1 day. It was cooled and filtered. The filtrate wasdiluted with EtOAc, washed with brine, dried and concentrated. The crudeproduct was purified on silica gel column with hexanes/CH₂Cl₂, 2:1 then1:1 to hexanes/EtOAc, 1:1 to give 12.7 g of desired product.

[1000] The MOM protection was carried out under the same conditions aspreviously described. After purification 12.7 g of product was obtained.

[1001] The mixture of chlorophenol (12.7 g), NaN₃ (6 g) in 20 mL of DMFwas heated at 70° C. overnight. After cooled, it was diluted with EtOAc,washed with water, brine, dried and concentrated. The crude product waspurified on silica gel colurnn to give 10.3 g of desired azide.

[1002] All above transformations were carried out under the conditionsidentical to previous series.

[1003] Preparation of ER804730:

[1004] ER804730 was prepared in the similar way as ER804104 from560-ycs-171.

[1005] Preparation of ER805135:

[1006] 2-Imidazole-carboxyaldehyde (96 mg, 1.0 mmol) in 0.5 mL of DMSOwas introduced to a mixture of 560-ycs-183 (0.30 g, 0.50 mmol) and 4 Amolecular sieves (0.30 g) in 5 mL of CH₂Cl₂. The mixture was stirred for30 minutes at rt after the addition of AcOH (28 μL, 0.50 mmol). K₂CO₃solid (0.14 g, 1.0 mmol), celite (0.5 g) and 10 mL of Et₂O were and themixture was filtered. The filtrate was concentrated before it wasdissolved in 5 ML of EtOH and treated with NaBH₄ at RT for 5 minutes.The mixture was then diluted with brine and EtOAc. Organic layer wasdried and concentrated to furnish 560-ycs-275 (0.27 g, 0.40 mmol) in 80%yield. MS: 676 (M⁺+1, 100%).

[1007] Transformations from 560-ycs-275 to 560-ycs-276 and 560-ycs-277to 560-ycs-279 were similar as the procedure for the preparation ofER804104.

[1008] Boc₂O (0.27 g, 1.2 mmol) was added to a solution of 560-ycs-276(0.29 g, 0.50 mmol) and Et₃N (0.21 mL, 1.5 mmol) in 5 mL of CH₂Cl₂. Themixture was stirred at rt for 12 hours before it was concentrated andpurified by silica gel chromatography to provide 560-ycs-277 (0.34 g,0.44 mmol) in 88% yield. MS: 794 (M⁺+Na, 100%).

[1009] A solution of 560-ycs-279 (0.22 g, 0.29 mmol) in 2 mL of TFA and2 mL of CH₂Cl₂ was sat at rt for 30 minutes before it was concentratedand dissolved in 2 mL of CH₃CN and 2 mL of 1 N HCl. The solution wasdiluted with 10:1 CHCl₃-MeOH after it was stirred at rt for 12 hours.Organic phase was washed with NaHCO₃, concentrated and purified bysilica gel chromatography to afford ER-805135 (60 mg, 0.12 mmol) in 25%overall yield for 6 steps.

[1010] Preparation of ER804744:

[1011] Ph₃P (35 mg, 0.14 mmol) was added to a solution of 560-ycs-171(28 mg, 0.045 mmol) in 3 mL of 2:1 THF-H₂O. The solution was stirred for12 hours at rt before all the volatile was evaporated and the cruderesidue was dissolved in 2 mL of CH₂Cl₂. Et₃N (44 μL, 0.32 mmol) andMsCl (17 μL, 0.22 mmol) was added at 0° C. The mixture was stirred at 0°C. for 10 minutes before it was washed with NaHCO₃ and concentrated toprovide 560-ycs-184 (21 mg, 0.031 mmol) in 69% for 2 steps.

[1012] ER804744 was prepared in the similar way as ER804101 from560-ycs-184.

[1013] Preparation of ER804759:

[1014] ER804759 was prepared in the similar way as ER804606 from560-ycs-36 and 2-(N,N-dimethylamino)-1-ethanol.

[1015] Preparation of 4 Carbon Linker C (14) Common Intermediate

[1016] 491-HAD-185I

[1017] Chlorobutan-1-ol (15.5 g, 143 mmol) and DEAD (28.8 g, 165 mmol)were added simultaneously over 45 min to a cooled (0° C. ice/water bath)solution of diphenolic toluide (20.0 g, 110 mmol) in THF (175 mL) andtoluene (700 mL) under a nitrogen atmosphere. Then, the ice/water bathwas removed and the reaction mixture was allowed to warm to rt. Afterstirred for 2.5 h, standard Mitsunobu workup conditions followed byflash chromatography (silica gel, 5-10% EtOAc/hexanes) yielded491-HAD-185 as a colorless solid (15.66 g, 52%)

[1018] 491-HAD-191

[1019] DBU (59.0 g, 388 mmol) was added drop wise to a cooled (salt/icebath) solution of 491-HAD-185 (14.4 g, 52.9 mmol) DMF (200 mL undernitrogen atmosphere followed by chloromethyl methyl ether (26.7 g, 332mmol). After stirred for 0.5 h, water (200 mL) was added and the aqueousphase was extracted with CH₂Cl₂. The combined CH₂Cl₂ extracts were driedwith anhydrous Na₂SO₄ and solvent was removed by reduced pressure. Flashchromatography (silica gel, 15% EtOAc/hexanes) gave 491-HAD-191 ascolorless oil (13.9 g, 81%).

[1020] 491-HAD-192

[1021] Sodium azide (8.64 g, 133 mmol) was added to a solution of491-HAD-191 in DMF (150 mL) and the suspension was heated to 80° C.After stirring 2 h water (200 mL) and CH₂Cl₂ (150 mL) was added. Theaqueous phase was extracted several times with CH₂Cl₂, dried withanhydrous Na₂SO₄ and solvent removed under reduced pressure. Flashchromatography (silica gel, 20% EtOAc/hexanes) afforded 491-HAD-192 aspale yellow oil (12.9 g, 90%).

[1022] 491-HAD-194

[1023] Lithium diisopropylamine was prepared under nitrogen atmospherein usual manner from diisopropylamine (12.8 mL, 91.5 mmol) andn-butyllithium in 5% HMPA/THF (150 mL) in flask equipped with overheadmechanical stirring. LDA solution was cooled to −78° C. (dry ice/acetonebath). Next, a solution of 491-HAD-192 in 5% HMPA/THF (35 mL) was added.After stirring 20 min, a solution of diphenyl diselenide (12.4 g, 39.7mmol) in 5% HMPA/THF (35 mL) was added. Small amount of intermediate wasobserved and additional solvent (20 mL) was added in an attempt tocreate a homogeneous solution. After stirred for 2.5 h, aqueous NH₄Clwas added and the aqueous phase was extracted several times with EtOAc.Combined organic extracts were washed with brine, dried with anhydrousNa₂SO₄ and solvent was evaporated under reduced pressure. Slightlyimpure 491-HAD-194 was obtained as a yellow oil (3.95 g, 21%) followingflash chromatography (silica gel, 20-100% hexanes/CH₂Cl₂, 2%EtOAC/CH₂Cl₂).

[1024] 491-HAD-196

[1025] A solution of 491-HAD-194 (3.94 g, 8.24 mmol) in ethanol (200proof, 24.5 mL) and 2.5 N NaOH (16.5 mL, 41.3 mmol) was heated to 60° C.After stirred for 2 days, the reaction solution was then diluted withwater and hexanes and the aqueous layer was acidified with NaHSO₃ andextracted with EtOAc. The combined organic extracts were dried withanhydrous Na₂SO₄ and evaporation of the solvent under reduced pressureto afford 491-HAD-196 (3.71 g) as a pale yellow solid. 491-HAD-196 wasnot purified and was put directly into next step.

[1026] 491-HAD-200

[1027] A flask equipped with nitrogen inlet was charged with 491-HAD-196(3.70 g, 7.97 mmol), CH₂Cl₂ (56 mL), DCC (6.58 g, 31.9 mmol), DMAP (97.3mg, 0.797 mmol) and triethylamine (4.44 mL, 31.9 mmol). After stirringseveral minutes 2-(trimethylsilyl) ethanol was added and the reactionmixture was heated to 35° C. for 3 days then stirred an additional 10-15h at rt. Toluene (100 mL) was added and the reaction mixture wasfiltered. The filtrate was washed with saturated aqueous NaHCO₃ solutionfollowed by brine, dried with anhydrous Na₂SO₄ and solvent was removedunder reduced pressure. Flash chromatography (silica gel, 20%EtOAc/hexanes) afforded 491-HAD-200 (1.89 g, 42%) as a colorless oil.

[1028] 491-HAD-230

[1029] A solution containing iodide 554-RB-260 (1.04 g, 1.72 mmol) and491-HAD-200 (2.53 mmol) in 5% HMPA/THF (2.53 mL), under nitrogenatmosphere, was cooled to −78° C. (dry ice/acetone bath) and thereaction vessel was shielded from light. 1M LiHMDS in THF (2.53 mL, 2.53mmol) was added over 75 min by syringe pump. After stirred for anadditional 40 min at −78° C., aqueous NH₄Cl was added and the aqueousphase was extracted several times with EtOAc. Combined organic extractswere washed with brine, dried with anhydrous Na₂SO₄ and solventevaporated under reduced pressure. Flash chromatography (silica gel,5-10-15% EtOAc/hexanes) gave slightly impure (small amount of491-HAD-200) 491-HAD-230 (1.86 g) of as a colorless oil.

[1030] 491-HAD-232

[1031] A solution of 491-HAD-230 (1.89 g, 1.82 mmol) in CH₂Cl₂ (26 mL)under a nitrogen atmosphere, was cooled to 0° C. (ice/water bath). Nextm-CPBA (57% 1.65 g, 5.46 mmol) was added in one portion. After stirredfor 1.5 h, triethylamine was added and ice/water bath was removed. Afterstirred for an addition hour at rt, the reaction mixture was cooled withice/water bath and a solution composed of 10% v/v Na₂S₂O₃ (aqueous,saturated)/NaHCO₃ (aqueous, saturated) was added. The aqueous phase wasextracted several times with CH₂Cl₂. Combined CH₂Cl₂ extracts werewashed with saturated NaHCO₃, dried with anhydrous Na₂SO₄ and solventwas evaporated under reduced pressure. Flash chromatography of theresidue (silica gel, 15% EtOAc/hexanes) gave 491-HAD-232 as pale yellowoil (0.96 g 63% over two steps).

[1032] 491-HAD-235

[1033] To a solution of 491-HAD-232 (0.96 g, 1.1 mmol) in THF (5 mL),under nitrogen atmosphere, a TBAF (1M/THF) solution (5.45 mL, 5.45mmol), buffered with imidazole HCl (0.14 g, 1.3 mmol) was added and thereaction solution was allowed to stir at rt. After stirred for 4 days,additional TBAF (1M in THF) (2.2 mL, 2.2 mmol) was introduced to thereaction flask and heating was increased to 50° C. After stirred for 15h, heating was stopped and cooled to rt, a saturated aqueous solution ofNH₄Cl was added. The aqueous phase was extracted several times withEtOAc. Combined organic phase was washed with brine, dried withanhydrous Na₂SO₄ and solvent was removed under reduced pressure. Thecrude residue was used for next step without purification.

[1034] 491-HAD-237

[1035] To a solution containing 2-chloro-1-methylpyridium iodide (1.65g, 6.47 mmol), CH₂Cl₂ (80 mL) and tributylamine (1.54 mL, 6.47 mmol),heated to reflux under a nitrogen atmosphere, a solution of crude491-HAD-235 in dichloromethane (160 mL) was added over 3.5 h usingsyringe pump. After stirred for an additional hour, the heat was shutoff and the reaction was then allowed to stir at rt overnight. Thereaction solution then was concentrated under reduced pressure and theresidue was diluted with EtOAc and water. The aqueous phase wasextracted several times with EtOAc and the combined organic phase waswashed with 0.05 M HCl (3×85 mL), saturated aqueous NaHCO₃. brinesuccessively, dried with Na₂SO₄ and concentrated down under reducedpressure. Flash chromatography of the residue (silica gel, 30% ethylacetate) afforded 491-had-237 as a pale yellow gel (0.46 g, 65% over twosteps).

[1036] Preparation of Imidazole C.14 Analogue, ER805023:

[1037] 491-HAD-251

[1038] A solution of 491-HAD-237 (71.4 mg, 0.110 mmol),triphenylphosphine (86.5 mg, 0.330 mmol) in THF (2 mL) and water (1 mL)was stirred at rt. After stirred for approximately 17 h, the reactionsolution was concentrated under reduced pressure. The residue wasazeotroped several times with toluene and used directly in the nextreaction without purification.

[1039] 491-HAD-252

[1040] To a flask charged with 491-HAD-254 (0.110 mmol) and molecularsieves (4A) in CH₂Cl₂, 2-imidazolecarboxaldehyde (21.1 mg, 0.220 mmol)in hot DMSO (0.30 mL) was added followed by acetic acid (6.3 μL). Afterstirred for 45 min at rt, anhydrous K₂CO₃ (0.030 g) was added. Thereaction mixture was subsequently diluted with diethyl ether andfiltered. Following the removal of solvent, the residue was re-dissolvedin methanol and resulting solution cooled with ice/water bath and NaBH₄(0.020 g, 0.53 mmol) was added. After stirred for 20 min, brine solutionwas added and the aqueous phase was extracted several times with EtOAc.Combined EtOAc extracts were dried with anhydrous Na₂SO₄ andconcentrated. The crude residue was not purified and was used directlyin next reaction.

[1041] 491-HAD-254

[1042] A solution containing 491-HAD-252 (0.110 mmol), 1 N NaOH (3.30mL, 3.30 mmol), ethanol (3.3 mL) and THF (1.6 mL) was stirred at 40-45°C. After 16 h, water was added followed by CH₂Cl₂. The aqueous phase wasextracted several times with CH₂Cl₂ and the combined organic phases weredried with anhydrous Na₂SO₄ and concentrated. The crude residue was notpurified and was used directly in next reaction.

[1043] 491-HAD-256

[1044] A solution of 491-HAD-254 (0.110 mmol), triethylamine (46.0 μL,0.330 mmol) and Boc anhydride (60.0 mg, 0.275 mmol) in CH₂Cl₂was stirredunder a nitrogen atmosphere at rt for 4 days. Reaction mixture wasconcentrated under reduced pressure. Flash chromatography (solventgradient used: 20% EtOAc/hexanes, 30% EtOAc/hexanes, 50% EtOAc/hexanes,75% EtOAc/hexanes) yielded 491-HAD-256 as a colorless gel (61.2 mg 70%over 4 steps).

[1045] 491-HAD-260

[1046] Reaction mixture containing 491-HAD-256 (59.1 mg, 0.0739 mmol),molecular sieves (4 47.8 mg), Celite (47.8 mg) and PCC (47.8 mg, 0.222mmol) in CH₂Cl₂ (1.70 mL) was stirred under a nitrogen atmosphere at rtfor 1 hr. Triethylamine (30.8 μL, 0.222 mmol) was added and reactionmixture was stirred for an additional 15 min. Diethyl ether was addedand reaction mixture was filtered through Celite. Volume of the filtratewas reduced and flash chromatography (100% EtOAc) afforded slightlyimpure 491-HAD-260 as a colorless oil (90.9 mg).

[1047] 491-HAD-261

[1048] To a solution of 491-HAD-260 in CH₂Cl₂ (0.60 mL), under anitrogen atmosphere, TFA was added. After stirring 30 min at rt, solventand volatiles were removed by rotary evaporation. Acetonitrile (0.6 mL)and 1 N HCl (0.60 mL, 0.60 mmol) were added to the residue. Afterstirring 19 h at rt, reaction mixture was cooled with ice/water bath anda saturated aqueous solution of NaHCO₃ was added. Aqueous phase wasextracted several times with CHCl₃ then once with 10% methanol/CHCl₃.Combined CHCl₃ extracts were dried with anhydrous Na₂SO₄ andconcentrated by reduced pressure. Flash chromatography (MeOH: CH₂Cl₂: 2MNH₃/MeOH 5:95:1, 10:90:1, 15:85:1) yielded 491-HAD-261, ER805023 as anoff-white solid (6.8 mg, 18% over 3 steps).

[1049] Preparation of Compound ER-804446 (C14 Difluoromethoxy)

[1050] Step 1

[1051] To a solution of compound 557-MS-262 (4.14 g, 6.69 mmol) in THF(40 mL) was added a 1M solution of TBAF (6.69 mL; 6.69 mmol). Thereaction mixture was stirred at room temperature for 30 minutes thenworked up in the usual manner. Chromatographic purification gavecompound 557-MS-151 (2.34 g, 92%).

[1052] Step 2

[1053] To a vigorously stirred solution of compound 557-MS-151 (520 mg,1.36 mmol) in dioxane (2 mL) at 55-60° C., was added pre-heated (50° C.)40% aqueous NaOH solution (2 mL). A stream of chlorodifluoromethane gaswas then admitted continuously to the reaction mixture via a gas inlettube (the tip of which was positioned just below the surface of thereaction mixture). After 25 minutes the reaction mixture was worked upin the usual manner. Chromatographic purification gave compound557-MS-154 (329 mg, 56%).

[1054] Step 3

[1055] A solution of 557-MS-154 (455 mg, 1.05 mmol) in ethanol (5 mL)was treated with 40% aqueous NaOH solution (2 mL) and heated underreflux for 16 hours. The reaction mixture was cooled to roomtemperature, diluted with water and then washed with diethyl ether. Theaqueous phase was acidified (with cooling) to pH3 by drop wise additionof concentrated aqueous hydrochloric acid. Extraction with diethyl ether(×4) followed by drying etc gave compound 557-MS-158 (384 mg, 88%).

[1056] Step 4

[1057] A solution of compound 557-MS-158 (384 mg, 0.92 mmol) in diethylether (8 mL) was treated with toluene (2 mL), triphenylphosphine (290mg, 1.10 mmol), and 2-(trimethylsilyl) ethanol (0.172 ml, 1.20 mmol),then cooled to 0° C. under an inert atmosphere. Diethylazidodicarboxylate (0.174 ml, 1.10 mmol) was added drop wise and thereaction mixture then allowed to warm to room temperature. After 3 hoursthe reaction mixture was worked up in the usual manner. Chromatographicpurification gave compound 557-MS-165 (410 mg, 86%).

[1058] Step 5

[1059] A mixture of compound 557-MS-165 (410 mg, 0.79 mmol) and compound554-RB-260 (523 mg, 0.72 mmol) was dissolved in THF (3.2 mL), treatedwith HMPA (0.6 mL) and then cooled to −78° C. under an inert atmosphere.A 0.5M solution of LiHMDS in THF (1.73 mL, 0.864 mmol) was then addeddrop wise over approximately 15 minutes. The reaction mixture wasstirred at −78° C. for 40 minutes, then warmed to 0° C. The intermediatecrude product was worked up in the usual manner and then dissolved indichloromethane (12 mL) and cooled to 0° C. A solution of approximately55% m-CPBA (452 mg) in dichloromethane (8 mL) was added portion wise.After 40 minutes triethylamine (1 mL) was added and the reaction mixturewas worked up in the usual manner. Chromatographic purification gavecompound 557-MS-203 (552 mg, 80%).

[1060] Step 6

[1061] A solution of compound 557-MS-203 (552 mg, 0.575 mmol) in THF(5.75 mL) was treated with a 1M solution of TBAF in THF (11.5 mL, 11.5mmol) then heated at 60° C. for approximately 3 hours. The usual work upgave crude compound 557-MS-205 (350 mg), which was used in the nextstage without purification.

[1062] Step 7

[1063] A solution of crude compound 557-MS-205 (assumed to contain 0.32mmol) in dichloroethane (66 mL) was added slowly to a heated solution(85° C.) of 2-chloro-1-methylpyridinium iodide (824 mg, 3.2 mmol) andtri-n-butylamine (0.768 mL, 3.2 mmol) in dichloroethane (100 mL). Thereaction mixture was heated at 85° C. for 1 hour post complete additionthen cooled to room temperature. The usual work up and chromatographicpurification gave compound 557-MS-212 (93 mg, 48% from compound557-MS-203).

[1064] Step 8

[1065] A solution of compound 557-MS-212 (93 mg, 0.15 mmol) in THF (6mL) and ethanol (12 mL) was treated with 1M aqueous NaOH solution (2.5mL) and heated at 60° C. for 1.5 hours, then at 70° C. for 1 hour. Theusual work up gave crude compound 557-MS-214 (74 mg), which was used inthe next stage without purification.

[1066] Step 9

[1067] A solution of crude compound 557-MS-214 (89 mg, 0.178 mmol) indichloromethane (10 mL) was treated with PCC (462 mg, 2.14 mmol) in thepresence of powdered 4 Å molecular sieves (462 mg). The reaction mixturewas stirred vigorously for 120 minutes at room temperature. Basificationwith excess triethylamine, followed by chromatographic purification gavecompound 557-MS-216 (53 mg, 60% from compound 557-MS-212).

[1068] Step 10

[1069] A solution of compound 557-MS-216 (53 mg, 0.106 mmol) in amixture of acetonitrile (7 mL) and dichloromethane (1.7 mL) was treatedwith 48% aqueous hydrofluoric acid (1.7 mL). After 35 minutes at roomtemperature the usual work up, followed by chromatographic purification,gave compound ER-804446 (40 mg, 91%) (m/z: 411.3 [M+1; 100%]).

[1070] Preparation of Compound ER-804387 (C14 Trifluoroethoxy)

[1071] Step 1

[1072] To a solution of 557-MS-151 (1 g, 2.62 mmol) in acetone (20 mL)were added potassium carbonate (440 mg, 3.14 mmol) and2,2,2-trifluoroethyl trichloromethanesulphonate (880 mg, 3.14 mmol). Thereaction mixture was heated at 70° C. for 2 hours then treated withfurther aliquots of potassium carbonate (440 mg; 3.14 mmol) and2,2,2-trifluoroethyl trichloromethanesulphonate (880 mg, 3.14 mmol).After a further 2 hours at 70° C. the reaction mixture was worked up inthe usual manner. Chromatographic purification gave compound 557-MS-161(760 mg, 63%).

[1073] Step 2

[1074] A solution of 557-MS-161 (760 mg, 1.64 mmol) in ethanol (5 mL)was treated with 40% aqueous NaOH solution (2 mL) and heated underreflux for 16 hours. The reaction mixture was cooled to roomtemperature, diluted with water and then washed with diethyl ether. Theaqueous phase was acidified (with cooling) to pH3 by drop wise additionof concentrated aqueous hydrochloric acid. Extraction with diethyl ether(×4) followed by drying etc gave compound 557-MS-163 (648 mg, 88%).

[1075] Step 3

[1076] A solution of compound 557-MS-163 (648 mg, 1.44 mmol) in diethylether (12 mL) was treated with toluene (3 mL), triphenylphosphine (454mg, 1.73 mmol), and 2-(trimethylsilyl) ethanol (0.269 mL, 1.875 mmol),then cooled to 0° C. under an inert atmosphere. Diethylazidodicarboxylate (0.272 mL, 1.73 mmol) was added drop wise and thereaction mixture then allowed to be warmed to room temperature. After1.5 hours the reaction mixture was worked up in the usual manner.Chromatographic purification gave compound 557-MS-167 (730 mg, 92%).

[1077] Step 4

[1078] A mixture of compound 557-MS-167 (730 mg, 1.33 mmol) and compound554-RB-260 (885 mg, 1.21 mmol) was dissolved in THF (9 mL), treated withHMPA (1 mL) and then cooled to −78° C. under an inert atmosphere. A 0.5Msolution of LiHMDS in THF (2.9 mL, 1.45 mmol) was then added drop wiseover approximately 20 minutes. The reaction mixture was stirred at −78°C. for 35 minutes, then warmed to 0° C. The intermediate crude productwas worked up in the usual manner and purified partially bychromatography. The intermediate was dissolved in dichloromethane (15mL) and cooled to 0° C. A solution of approximately 55%meta-chloroperbenzoic acid (612 mg) in dichloromethane (10 mL) was addedportion wise. After 30 minutes triethylamine (1.37 mL) was added and theusual work up, followed by chromatographic partial purification, gaveimpure compound 557-MS-177 (950 mg), which was used in the next stagewithout further purification.

[1079] Step 5

[1080] A solution of crude compound 557-MS-177 (475 mg, assumed tocontain 0.479 mmol) in a 1M solution of TBAF in THF (9.58 mL; 9.58 mmol)was heated at 50° C. for approximately 7 hours. The usual work up gavecrude compound 557-MS-179 (300 mg), which was used in the next stagewithout purification.

[1081] Step 6

[1082] A solution of crude compound 557-MS-179 (assumed to contain 0.153mmol) in dichloroethane (30 mL) was added slowly to a heated solution(85° C.) of 2-chloro-1-methylpyridinium iodide (391 mg, 1.53 mmol) andtri-n-butylamine (0.365 mL, 1.53 mmol) in dichloroethane (100 mL). Thereaction mixture was heated at 85° C. for 1 hour post complete additionthen cooled to room temperature. The usual work up and chromatographicpurification gave compound 557-MS-183 (40 mg, 41% from compound557-MS-167).

[1083] Step 7

[1084] A solution of compound 557-MS-183 (40 mg, 0.063 mmol) in THF (2.5mL) and ethanol (5 mL) was treated with 1M aqueous NaOH solution (1 mL)and heated at 60° C. for 3.5 hours. The usual work up gave crudecompound 557-MS-191 (32 mg), which was used in the next stage withoutpurification.

[1085] Step 8

[1086] A solution of crude compound 557-MS-191 (52 mg, 0.098 mmol) indichloromethane (6 mL) was treated with PCC (253 mg, 1.18 mmol) in thepresence of powdered 4 Å molecular sieves (253 mg). The reaction mixturewas stirred vigorously for 3 hours at room temperature. Basificationwith excess triethylamine, followed by chromatographic purification gavecompound 557-MS-194 (39.3 mg, 76% from compound 557-MS-183).

[1087] Step 9

[1088] A solution of compound 557-MS-194 (23 mg, 0.0435 mmol) in amixture of acetonitrile (3.2 mL) and dichloromethane (0.8 mL) wastreated with 48% aqueous hydrofluoric acid (0.8 mL). After 35 minutes atroom temperature the usual work up, followed by chromatographicpurification, gave compound ER-804387 (15.8 mg, 82%).

[1089] Preparation of Compound B2356 (C14 Hydroxy) & Compound B2359 (C14OMOM):

[1090] A solution of methyl 2,4-dihydroxy-6-methylbenzoate (6 g, 32.95mmol) in dry DMF (10 mL) was added to a well-stirred suspension ofhexane-washed sodium hydride (3.95 g, 60% in mineral oil; approximately98.85 mmol) in dry DMF (50 mL), at 0° C. under an inert atmosphere. Thereaction mixture was stirred at 0° C. for 30 minutes then treated withmethoxymethyl chloride (5.26 mL, 69.2 mmol) drop wise. The reactionmixture was stirred for 2 hours then worked up in the usual manner togive compound 453-MS-21 (8.14 g, 91%).

[1091] Step 2

[1092] To a solution of diisopropylamine (3.14 mL, 22.4 mmol) in dry THF(45 mL), at −20° C. under an inert atmosphere, was added drop wise a2.5M solution of n-butyllithium in hexanes (8.96 mL, 22.4 mmol). Thereaction mixture was allowed to warm to 0° C., stirred for 10 minutes at0° C., then cooled to −78° C. A solution of compound 453-MS-21 (4.03 g,14.9 mmol) in dry THF (15 mL) was added drop wise. The reaction mixturewas stirred at −78° C. for 1 hour then treated with a solution ofdiphenyl diselenide (5.59 g, 17.9 mmol) in dry THF (18 mL). The reactionmixture was stirred at −78° C. for 30 minutes then worked up in theusual manner. Chromatographic purification gave compound 453-MS-108(3.47 g, 54%).

[1093] Step 3

[1094] A solution of compound 453-MS-108 (2.16 g, 5.08 mmol) in ethanol(20 mL) was treated with powdered NaOH (610 mg, 15.24 mmol) and heatedunder reflux for 28 hours. The reaction mixture was cooled to roomtemperature and concentrated under reduced pressure (to a residualvolume of approximately 5 mL). The residue was partitioned between waterand diethyl ether. The aqueous fraction was acidified to pH3 by slowaddition of 1M aqueous HCl (approximately 16 mL), with cooling. Theacidic solution was extracted with diethyl ether, and the extracts thenwashed immediately with saturated aqueous brine solution (at least fivetimes). Drying etc gave compound 453-MS-110 (2.016 g, 97%).

[1095] Step 4

[1096] To a solution of compound 453-MS-110 (2.016 g, 4.9 mmol) indiethyl ether (40 mL), at 0° C. under an inert atmosphere, were addedtoluene (10 mL), triphenylphosphine (1.41 g, 5.39 mmol), and2-(trimethylsilyl) ethanol (0.843 mL, 5.88 mmol). Diethylazidodicarboxylate (0.849 mL, 5.39 mmol) was then added drop wise. Thereaction mixture was allowed to warm to room temperature and stirred forapproximately 16 hours. The usual work up followed by chromatographicpurification gave compound 453-MS-111 (2.24 g, 90%).

[1097] Step 5

[1098] A mixture of compound 453-MS-111 (1.33 g, 2.6 mmol) and compound343-YW-281 (866 mg, 1.46 mmol) was dissolved in a solution of 10% HMPAin THF (15 mL) and cooled to −78° C. under an inert atmosphere. A 1Msolution of LiHMDS in THF (2.19 mL, 2.19 mmol) was then added drop wiseover approximately 15 minutes. After another 45 minutes an extra aliquotof 1M solution of LiHMDS in THF was added (0.438 mL, 0.438 mmol). Thereaction mixture was warmed to 0° C. and the intermediate crude productworked up in the usual manner, and purified partially by chromatography.The intermediate was then dissolved in dichloromethane (14 mL) andcooled to 0° C. A solution of approximately 55% meta-chloroperbenzoicacid (280 mg) in dichloromethane (6 mL) was added. After 10 minutes,extra 55% m-CPBA (28 mg) was added. The reaction mixture was stirred at0° C. for a further 20 minutes, then treated with triethylamine (1.22mL) and worked up in the usual manner. Chromatographic purification gavecompound 453-MS-77 (625 mg, 52%).

[1099] Step 6

[1100] To a vigorously stirred biphasic mixture of compound 453-MS-77(618 mg, 0.753 mmol), dichloromethane (20 mL) and water (10 mL), wasadded DDQ (190 mg, 0.84 mmol). After 1 hour at room temperature thereaction mixture was worked up in the usual manner. Chromatographicpurification gave compound 453-MS-82 as a mixture of 4 diastereoisomers(381 mg, 72%).

[1101] Step 7

[1102] A solution of compound 453-MS-82 (381 mg, 0.544 mmol) in THF (10mL) was treated with TBAF (284 mg, 1.09 mmol) and stirred at roomtemperature for approximately 16 hours. The usual work up gave compound453-MS-84 (326 mg, quantitative), as a mixture of 4 diastereoisomers.

[1103] Step 8

[1104] To a solution of triphenylphosphine (109 mg, 0.417 mmol) in dryTHF (6 mL), at room temperature under an inert atmosphere, was addeddiethyl azidodicarboxylate (66 μL, 0.417 mmol) drop wise overapproximately 30 seconds. A solution of compound 453-MS-84 (167 mg,0.278 mmol) in dry THF (10 mL) was added drop wise over approximately 10minutes. The reaction mixture was stirred at room temperature for 10minutes then treated with extra triphenylphosphine (36 mg, 0.137 mmol),followed by extra diethyl azidodicarboxylate (22 μL, 0.137 mmol). Thereaction mixture was stirred for a further 10 minutes then worked up inthe usual manner. Partial chromatographic purification gave compound453-MS-91 (122 mg, 76%) as a mixture of 4 diastereoisomers.

[1105] Step 9

[1106] A solution of compound 453-MS-91 (70 mg, 0.12 mmol) in ethanol(2.5 mL) was treated with THF (1.25 mL) and 1M aqueous NaOH (0.6 mL, 0.6mmol) and stirred at room temperature for approximately 6 days.Chromatographic purification gave two fractions of partially resolveddiastereoisomers:

[1107] Fraction A (less polar): a mixture of 2 diastereoisomers—compound453-MS-101A (24 mg);

[1108] Fraction B (more polar): a mixture of 2 diastereoisomers—compound453-MS-101B (25 mg);

[1109] (total yield: 49 mg, 86%)

[1110] Step 10

[1111] A solution of compound 453-MS-101B (25 mg, 52.3 μmol) indichloromethane (1.5 mL) was treated with PCC (135 mg, 0.627 mmol) inthe presence of powdered 4 Å molecular sieves (135 mg). The reactionmixture was stirred vigorously for 40 minutes at room temperature.Basification with excess triethylamine, followed by chromatographicpurification gave compound 453-MS-116 (14 mg, 63%).

[1112] Step 11

[1113] A solution of compound 453-MS-116 (13 mg, 27.3 μmol) in a mixtureof dioxane (3 mL) and deuterium oxide (3 mL) was treated with Dowex®(50WX8- 100, 200 mg), and stirred at room temperature for approximately16 hours. The usual work up, followed by purification using reversedphase HPLC, gave compound B2356 (2.4 mg, 25%) [m/z: 349.3 (M+1, 60%),161.0 (100%)], and compound B2358 (2 mg, 20%).

[1114] Preparation of Compound B2357 (C14 Hydroxy) & Compound B2359 (C14OMOM):

[1115] Step 1

[1116] A solution of compound 453-MS-101A (24 mg, 50.1 μmol) indichloromethane (2 mL) was treated with pyridinium chlorochromate (130mg, 0.602 mmol) in the presence of powdered 4 Å molecular sieves (130mg). The reaction mixture was stirred vigorously for 1 hour at roomtemperature. Basification with excess triethylamine, followed bychromatographic purification gave compound 453-MS-122 (19 mg, 80%).

[1117] Step 2

[1118] A solution of compound 453-MS-122 (21 mg, 44 μmol) in a mixtureof acetonitrile (4.6 mL) and dichloromethane (1.1 mL) was treated with48% aqueous hydrofluoric acid (1.1 mL), and stirred at room temperaturefor approximately 2 hours. The usual work up, followed by purificationusing reversed phase HPLC, gave compound B2357 (3.5 mg, 23%) [m/z: 349.2(M+1, 50%), 161.0 (100%)], and compound B2359 (1.6 mg, 10%).

[1119] Preparation of C14-C, H, or Halogen Analogs, NF0887, NF2433,NF2435, NF2436, NF2557 and ER-805053,

[1120] Synthetic Procedure for NF2433

[1121] Using same procedure for 16, YE-06 (526 mg, 0.714 mmol) wasconverted to NY-78 (657 mg).

[1122] Using same procedure for 18, NY-78 (653 mg, 0.644 mmol) wasconverted to NY-79 (529 mg).

[1123] Using same procedure for 509-HD-116, NY-79 (528 mg, 0.584 mmol)was converted to NY-80 (231 mg). NY-80 was used without purification forthe next step.

[1124] Using same procedure for TM-12, NY-80 (230 mg, 0.511 mmol) wasconverted to NY-81 (157 mg).

[1125] Using same procedure for 509-HD-125, NY-81 (127 mg, 0.294 mmol)was converted to NY-82 (118 mg).

[1126] Using same procedure for NF-0675, NY-82 (118 mg, 0.274 mmol) wasconverted to NF-2433 (79 mg).

[1127] Synthetic Procedure for NF-2436

[1128] Using same procedure for 9, NY-83 (7.46 g, 40 mmol) was convertedto NY-84 (11.39 g).

[1129] To a mixture of NY-84 (3 g, 10 mmol), Et₂NH (2.07 mL, 20 mmol)and THF (80 mL), isopropylmagenesium chloride(2M in THF, 10 mL, 20 mmol)was gradually added at −30° C. The reaction mixture was allowed to warmto 0° C. The reaction mixture was quenched with sat. NaHCO₃ andextracted with EtOAc. The organic layer was washed with water, brine,dried over Na₂SO₄, filtered and concentrated. The crude product waspurified on silica gel column with hexane/EtOAc, 10:1, 8:1, 6:1 to give3.21 g of NY-85.

[1130] NY-85 (3.2 g, 9.36 mmol) and TMEDA (2.2 mL, 14.58 mmol) weredissolved in THF (30 mL) and cooled to −78° C., under nitrogen. Then,sec-BuLi (1.3M/cyclohexane, 11 mL, 14.3 mmol) was slowly added and thereaction was stirred at −78° C. for 1 hr. DMF (1.4 mL, 18.08 mmol) wasadded to the solution, then the solution was stirred at −78° C. for 30min. The mixture was quenched with sat.NH₄Cl and extracted with EtOAc.The organic layer was washed with water, brine and dried over Na₂SO₄,filtered and concentrated. The crude product was purified on silica gelcolumn with hexane/EtOAc, 6:1 to give 2.56 g of NY-86.

[1131] To a solution of NY-86 (2.55 g, 6.89 mmol) in MeOH (30 mL), NaBH₄(260 mg, 6.87 mmol) was added at 0° C. and stirred for 30 min. Thereaction mixture was quenched with sat. NH₄Cl and extracted with EtOAc.The organic layer was washed with water, brine, dried over Na₂SO₄,filtered and concentrated to give 2.5 g of NY-87.

[1132] A mixture of NY-87 (2.5 g, 6.72 mmol), AcOH (1.92 mL, 33.54 mmol)and EtOH (50 mL) was refluxed for 4 hrs. The mixture was concentratedand the residue was alkalized with

[1133] sat.NaHCO₃ and extracted with EtOAc. The aqueous layer wasreextracted with EtOAc (×2) and the oranic layers were washed with sat.NH₄Cl, water, brine, dried over Na₂SO₄, filtered and concentrated. Thecrude crystallized product was washed with Et₂O to give 718 mg of NY-88.The mother liquid was purified on silica gel column with hexane/EtOAc,3:1, 2:1 to give additional 230 mg of NY-88.

[1134] Using same procedure for 509-HD-209, NY-88 (940 mg, 5.09 mmol)was

[1135] To a solution of NY-89 (1.74 g, 7.61 mmol) in DMSO (20 mL), asolution of KOH (450 mg, 8.02 mmol) in water (10 mL) was added andstirred at room temperature for 30 min. Then, the mixture was stirred at40° C. for 30 min. MeI (9.5 mL, 153 mmol) was added to the reactionmixture, and the mixture was stirred at room temperature for 3 hrs. Thereaction mixture was quenched with sat. NH₄Cl and extracted with EtOAc.The organic layer was washed with water, 5% citric acid, water,sat.NaHCO₃, brine, dried over Na₂SO₄, filtered and concentrated to give19.3 g of NY-90

[1136] To a solution of NY-90 (1.93 g, 7.61 mmol), Ph2S2 (5 g, 22.9mmol), pyridine (3.7 mL, 45.75 mmol), ^(n)Bu₃P (5.7 mL, 22.88 mmol) wereadded at 0° C. The mixture was allowed to warm to room temperature andstirred for 12 hrs. The reaction mixture was diluted with EtOAc andwashed with 5% citric acid(×2), water, sat. NaHCO₃, brine, dried overNa₂SO₄, filtered and concentrated. The crude product was purified onsilica gel column with hexane/EtOAc, 15:1, 5:1 to give 1.9 g of NY-91.

[1137] Using same procedure for 509-HD-212, NY-91 (2.12 g, 6 mmol) wasconverted to NY-92 (2.07 g). NY-92 was used without purification for thenext step.

[1138] Using same procedure for 509-HD-213, NY-92 (2.07 g, 6 mmol) wasconverted to NY-93 (2.19 g).

[1139] Using same procedure for 16, YE-06 (505 mg, 0.685 mmol) wasconverted to NY-94 (625 mg).

[1140] Using same procedure for 18, NY-94 (623 mg, 0.594 mmol) wasconverted to NY-95 (501 mg).

[1141] Using same procedure for 509-HD-116, NY-95 (500 mg, 0.533 mmol)was converted to NY-96 (400 mg). NY-96 was used without purification forthe next step.

[1142] Using same procedure for TM-12, NY-96 (400 mg, 0.533 mmol) wasconverted to NY-97 (142 mg).

[1143] Using same procedure for 509-HD-125, NY-97 (139 mg, 0.298 mmol)was converted to NY-98 (120 mg).

[1144] Using same procedure for NF-0675, NY-98 (118 mg, 0.254 mmol) wasconverted to NF-2436 (24 mg).

[1145] Synthetic Procedure for NF2435

[1146] Using similar procedure for the synthesis of 509-HD-209 from509-HD-207, 3,5-dimethylphenol (27.58 g, 225.75 mmol) was converted tocolorless oil of MK-091 (15.91 g, 42%) as compound purified.

[1147] MK-091 (6.00 g, 36.10 mmol) was dissolved in 18 mL of dry Et₂O.To the stirred solution was added n-BuLi in hexane (1.6M, 27 mL, 43.32mmol, 1.2 eq.) and the mixture was warmed to 40 ° C. After 30 min at 40° C. the color of the reaction mixture turned dark red. The mixture wascooled to −78 ° C. and excessive dry CO₂ gas (ca 30 eq.) was added bybubbling through an inlet over 30 min. Then, the resulting mixture wasallowed to warm to rt. After 30 min the reaction mixture was quenchedwith water and washed with Et₂O. The basic aqueous layer was acidifiedwith aqueous solution of KHSO₄ and extracted with AcOEt. The organicextract was washed with brine, dried over Na₂SO₄, filtered, andconcentrated to yield crude colorless crystals of MK-092 (4.55 g, <60%).The crude MK-092 was used for next step without purification.

[1148] Crude MK-092 (4.55 g, assumed to contain 21.6 mmol) was dissolvedin 200 mL of CH₃CN. To the solution were added CsCO₃ (5.64 g, 17.3 mmol)and MeI (2.20 mL, 34.6 mmol) and the mixture was stirred at rtovernight. After concentration of the reaction mixture, water was addedand extracted with AcOEt. The organic extract was washed with saturatedaqueous NaHCO₃ and brine, then dried over Na₂SO₄, filtered, andconcentrated to give crude oil of MK-093. The crude product was purifiedby chromatography on silica gel (hexane/AcOEt: 10/1) to afford paleyellow oil of MK-093 (3.19 g, 39% 2 steps).

[1149] Using similar procedure for the synthesis of 509-HD-211 from509-HD-209, MK-093 (2.86 g, 12.77 mmol) was converted to a mixture ofMK-094, MK-095 and MK-093. The crude product was purified bychromatography on silica gel (hexane/AcOEt: 9/1) to afford pale yellowoil of MK-094 (652 mg, 13%), the structure of which was determined byNOESY analysis.

[1150] Using similar procedure for the synthesis of 509-HD-212 from509-HD-211, MK-094 (650 mg, 1.71 mmol) was converted to crude benzoicacid.

[1151] Using similar procedure for the synthesis of MK-047 from MK-046,the crude benzoic acid was converted to pale pink oil of MK-096 (746 mg,94% 2 steps) as compound purified.

[1152] Using similar procedure for the synthesis of compound 5 fromcompound 2 and compound 3, YE-06 (516 mg, 0.700 mmol) coupled withMK-096 (489 mg, 1.05 mmol) was converted to colorless oil of MK-097 (491mg, 76% 3 steps) as compound purified.

[1153] Using similar procedure for the synthesis of YE-17 from YE-16,MK-097 (491 mg, 0.535 mmol) was converted to crude oil of MK-098 (486mg, including silyl impurity). The crude MK-098 was used for next stepwithout purification.

[1154] Using similar procedure for the synthesis of YE-18 from YE-17,the crude MK-098 (486 mg, assumed to contain 0.535 mmol) was convertedto colorless oil of MK-099 (159 mg, 67% 3 steps) as compound purified.

[1155] Using similar procedure for the synthesis of 509-HD-125 from509-HD-119B, MK-099 (158 mg, 0.354 mmol) was converted to crude paleyellow solid of MK-100 (146 mg, <93%). The crude MK-100 was used fornext step without purification.

[1156] Using similar procedure for the synthesis of NF1226 from MK-035,crude MK-100 (146 mg) was converted to crude pale yellow crystals. Thecrude product was purified by chromatography on silica gel(hexane/AcOEt: 3/2) to afford colorless crystals of NF2435 (88 mg, 69% 2steps).

[1157] Synthetic Procedure for NF2557, NF2558, and NF2559

[1158] Using similar procedure for the synthesis of 509-HD-209 from509-HD-207, TM-34 (12.23 g, 42.71 mmol) was converted to colorless oilof MK-120 (13.40 g, 95%) as compound purified.

[1159] MK-120 (13.38 g, 40.50 mmol) was dissolved in 200 mL of EtOH. 10%Pd on carbon (50% wet, 2.7 g) was added. The mixture was stirred underhydrogen at rt. After 8 hrs catalyst was filtered through Celite and thefiltrate was concentrated to give crude solid. The crude product waspurified by chromatography on silica gel (hexane/AcOEt: 2/1) to affordcolorless crystals of MK-121 (8.67 g, 89%).

[1160] Using similar procedure for the synthesis of 611-MS-88 from611-MS-84, MK-121 (8.66 g, 36.04 mmol) was converted to colorlesscrystals of MK-122 (13.20 g, 98%) as compound purified.

[1161] Using similar procedure for the synthesis of MK-073 from MK-072,propargyl alcohol (10.74 g, 191.6 mmol) was converted to colorless oilof MK-123 (27.03 g, 96%) as compound purified.

[1162] MK-122 (13.20 g, 35.46 mmol) and MK-123 (12.50 g, 70.92 mmol, 2.0eq.) were dissolved in 440 mL of DMF. To the solution were added Ph₃P(5.58 g, 21.28 mmol, 0.6 eq.), Pd(Ph₃P)₄ (6.15 g, 5.32 mmol, 0.15 eq.),CuI (1.01 g, 5.32 mmol, 0.15 eq.), and Et₃N (19.8 mL, 141.83 mmol, 4eq.). The mixture was heated to 45 ° C. and stirred for 1.5 hrs undernitrogen atmosphere. The mixture was cooled to rt and diluted withEt₂O-hexane, then stirred for a while. The resulting mixture wasfiltered through a pad of Celite to remove insoluble solid. The filtratewas washed with saturated aqueous solution of NH₄Cl and brine, driedover Na₂SO₄, filtered, and concentrated to yield crude product. Thecrude product was purified by chromatography on silica gel(hexane/AcOEt: 7/1 to 5/1) to afford pale brown oil of MK-124 (12.11 g,86%).

[1163] Using similar procedure for the synthesis of MK-074 from MK-073,MK-124 (14.28 g, 35.84 mmol) was converted to crude yellow oil of MK-125(14.34 g). The crude MK-125 was used for next step without purification.

[1164] Using similar procedure for the synthesis of MK-114 from MK-113,crude MK-125 (14.34 g, assumed to contain 35.84 mmol) was converted tocrude yellow oil of benzoic acid (14.92 g). The crude benzoic acid wasused for next step without purification.

[1165] Using similar procedure for the synthesis of MK-093 from MK-092,the crude benzoic acid (14.92 g, assumed to contain 35.84 mmol) wasconverted to colorless oil of MK-126 (10.92 g, 78% 3 steps) as compoundpurified.

[1166] Using modified procedure for the synthesis of 509-HD-211 from509-HD-209, MK-126 (960 mg, 2.47 mmol) was converted to crude selenideproduct. The crude product was purified by chromatography on silica gel(hexane/AcOEt: 6/1 to 5/1) to afford pale yellow oil (953 mg) as aninseparable mixture of desirable MK-127 (ca 710 mg, ca 60%), MK-128 (ca10%), and MK-126 (ca 17%). The mixture was used for next step withoutfurther purification.

[1167] Using similar procedure for the synthesis of MK-114 from MK-113,the mixture (953 mg) of MK-127 (ca 710 mg, assumed to contain 1.31mmol), MK-128, and MK-126 was converted to crude oil of benzoic acids(980 mg). The crude benzoic acids were used for next step withoutpurification.

[1168] Using similar procedure for the synthesis of MK-047 from MK-046,the crude mixture of benzoic acids (980 mg) was converted to crudeTMS-ethyl esters. The crude product was purified by chromatography onsilica gel (hexane/AcOEt: 6/1) to afford colorless oil (875 mg) as aninseparable mixture of desirable MK-129 (ca 683 mg, ca 83%) and otherTMS-ethyl esters corresponding to MK-128 and MK-126. The mixture wasused for next step without further purification.

[1169] Using similar procedure for the synthesis of compound 5 fromcompound 2 and compound 3, YE-06 (483 mg, 0.655 mmol) coupled with themixture (875 mg) including MK-129 (ca 683 mg, 1.08 mmol) was convertedto colorless oil of MK-130 (519 mg, 73% 3 steps).

[1170] Using similar procedure for the synthesis of YE-17 from YE-16,MK-130 (519 mg, 0.480 mmol) was converted to crude oil of MK-131 (620mg, including silyl impurity). The crude product was used for next stepwithout purification.

[1171] Using similar procedure for the synthesis of YE-18 from YE-17,the crude MK-131 (620 mg, assumed to contain 0.480 mmol) was convertedto crude oil of lactonized product. The crude product was purified bychromatography on silica gel (hexane/AcOEt: 5/2) to afford colorless oilof MK-132 (88 mg, 30% 3 steps) and colorless oil of des-MOM form MK-133(47 mg, 17% 3 steps) respectively.

[1172] Using similar procedure for the synthesis of 509-HD-125 from509-HD-119B, MK-133 (46 mg, 0.0812 mmol) was converted to crude paleyellow oil of enone (35 mg, <76%). The crude enone was used for nextstep without purification.

[1173] Using similar procedure for the synthesis of NF0531 from NF0530,the crude enone (35 mg, assumed to contain 0.0620 mmol) was converted tocolorless crystals of MK-134 (20 mg, 55% 2 steps).

[1174] Using similar procedure for the synthesis of NF0675 from TM-13,MK-134 (7 mg, 0.0157 mmol) was converted to colorless crystals of NF2557(5.1 mg, 80%) and colorless solid of NF2558 (1.5 mg, 19%) as compoundpurified, respectively.

[1175] Synthesis of ER-805053

[1176] Step 1

[1177] To a solution of methyl 2,4-dihydroxy-6-methylbenzoate (10.9 g,0.0598 mol) in dry N,N-dimethylformamide (100 mL) were added imidazole(4.48 g, 0.0658 mol) and tert-butyldiphenylsilyl chloride (15.6 mL,0.0658 mol). The reaction mixture was stirred at room temperature for 24hours then worked up in the usual manner. The crude product was purifiedchromatographically to give compound 557-MS-232 (12.33 g, 49%).

[1178] Step 2

[1179] To a solution of compound 557-MS-232 (9.08 g, 0.021 mol) in drytetrahydrofuran (100 mL), at 0° C. under an inert atmosphere, was addedsodium hydride (55% dispersion in oil; 1.88 g, approximately 0.042 mol).The reaction mixture was stirred at 0° C. for 30 minutes then treatedwith methoxymethyl chloride (3.28 mL, 0.042 mol). The reaction mixturewas allowed to warm to room temperature overnight. The usual work upgave compound 557-MS-233 (9.17 g, 91%).

[1180] Step 3

[1181] To a solution of freshly prepared lithium diisopropylamide (16.2mmol) in dry tetrahydrofuran (15 mL), at −78° C. under an inertatmosphere, was added drop wise a solution of compound 557-MS-233 (3.93g, 9.01 mmol) in dry tetrahydrofuran (15 mL). The reaction mixture wasstirred at −78° C. for 45 minutes then a solution of diphenyl diselenide(2.53 g, 8.11 mmol) in dry tetrahydrofuran (15 mL) was added rapidly(down the inside walls of the reaction vessel so as to pre-chill thediphenyl diselenide solution). The reaction mixture was stirred at −78°C. for 45 minutes then treated drop wise with a 2M solution of aceticacid in diethyl ether (8 mL). The reaction mixture was then worked up inthe usual manner. Chromatographic purification gave compound 557-MS-262(4.57 g, 82%).

[1182] Step 4

[1183] A solution of compound 557-MS-262 (6.29 g, 0.01 mol) in ethanol(100 mL) was treated with powdered sodium hydroxide (4 g, 0.1 mol) andheated under reflux for 30 minutes. The reaction mixture was cooled to5° C. and acidified to pH6.5 with 1M HCl. The majority of the ethanolwas then removed by concentration in vacuo and resultant residuepartitioned between water and ethyl acetate. The organic layer waswashed sequentially with saturated aqueous sodium bicarbonate solutionand water. Drying etc gave a crude residue, which was purifiedchromatographically to give compound 611-MS-84 (3.3 g, 87%).

[1184] Step 5

[1185] A solution of compound 611-MS-84 (1.65 g, 4.33 mmol) indichloromethane (20 mL), at 0° C., was treated sequentially withpyridine (0.385 mL, 4.76 mmol) and trimethylsulfonic anhydride (0.764mL, 4.54 mmol). After 25 minutes at 0° C. the reaction mixture waswarmed to room temperature and worked up in the usual manner to givecompound 611-MS-88 (1.5 g, 68%).

[1186] Step 6

[1187] A solution of compound 611-MS-88 (1.5 g, 2.92 mmol) in1,2-dimethoxyethane (25 mL) was heated under reflux, under an inertatmosphere, in the presence of phenyl boronic acid (713 mg, 5.84 mmol),palladium tetrakistriphenylphosphine (335 mg, 0.29 mmol), lithiumchloride (247 mg, 5.84 mmol) and 2M aqueous sodium carbonate (25 mL).After 2 hours the reaction mixture was cooled to room temperature. Theusual work up, followed by chromatographic purification gave compound611-MS-91 (1.004 g, 78%).

[1188] Step 7

[1189] A mixture of compound 611-MS-91 (512 mg, 1.16 mmol) and compound554-RB-260 (635 mg, 1.05 mmol) was dissolved in a solution of 10%hexamethylphosphoramide in tetrahydrofuran (8.8 mL) and cooled to −78°C. under an inert atmosphere. A 0.5M solution of lithiumbis-(trimethylsilyl) amide in tetrahydrofuran (2.52 mL, 1.26 mmol) wasthen added drop wise over approximately 30 minutes. The reaction mixturewas stirred at −78° C. for 2 hours, then warmed to 0° C. Theintermediate crude product was worked up in the usual manner and thendissolved in dichloromethane (15 mL) and cooled to 0° C. A solution ofapproximately 55% meta-chloroperbenzoic acid (724 mg) in dichloromethane(12 mL) was added. After 30 minutes triethylamine (1.6 mL) was added andthe reaction mixture was worked up in the usual manner. Chromatographicpurification gave compound 611-MS-102 (560 mg, 64%).

[1190] Step 8

[1191] A solution of compound 611-MS-102 (560 mg, 0.738 mmol) intetrahydrofuran (5 mL) was treated with a 1M solution oftetrabutylammonium fluoride in tetrahydrofuran (0.74 mL, 0.74 mmol). Theusual work up, followed by chromatographic purification gave compound611-MS-104 (340 mg, 71%).

[1192] Step 9

[1193] A solution of compound 611-MS-104 (340 mg, 0.527 mmol) in ethanol(10 mL) was treated with powdered sodium hydroxide (211 mg, 5.27 mmol)and heated under reflux. Cooling and acidification to pH6.5, followedthe usual work up gave crude compound 611-MS-106, which was used in thenext stage without purification.

[1194] Step 10

[1195] A solution of crude compound 611-MS-106 (assumed to contain 0.176mmol) in dichloromethane (20 mL) was added slowly to a heated solution(40° C.) of 2-chloro-1-methylpyridinium iodide (449 mg, 1.76 mmol) andtri-n-butylamine (0.42 mL, 1.76 mmol) in dichloromethane (60 mL). Theusual work up and chromatographic purification gave compound 611-MS-108(3 mg, 3% from compound 611-MS-104).

[1196] Step 11

[1197] A solution of compound 611-MS-108 (3 mg, 5.9 μmol) indichloromethane (500 μL) was treated with pyridinium chlorochromate (20mg, 88 μmol) in the presence of powdered 4 Å molecular sieves (20 mg).The reaction mixture was stirred vigorously for 4 hours at roomtemperature. Basification with excess triethylamine, followed bychromatographic purification gave compound 611-MS-118 (1.4 mg, 48%).

[1198] Step 12

[1199] A solution of compound 611-MS-118 (1.4 mg, 2.76 μmol) in amixture of acetonitrile (400 μL) and dichloromethane (100 μL) wastreated with 48% aqueous hydrofluoric acid (100 μL), and stirred at roomtemperature for 30 minutes. The usual work up followed bychromatographic purification gave compound ER-805053 (1.0 mg; 83%).

[1200] Preparation of C14-Aniline Analogs: ER805940, and ER806201:

[1201] Preparation of ER805940:

[1202] Tf₂O (0.42 ML, 2.5 mmole) was added to a solution of ER-805102(0.95 g, 1.7 mmole) and Et₃N (0.58 ML, 4.2 mmole) in 20 ML of CH₂Cl₂ at0° C. The mixture was stirred for 10 min before the addition of aq.NaHCO₃. Aq. layer was extracted twice with CH₂Cl₂. The organics wereconcentrated and passed through a short plug of silica gel (20%EtOAc/Hex).

[1203] The triflate thus obtained was added Pd(OAc)₂ (19 mg, 0.08mmole), BINAP (64 mg, 0.10 mmole) and Cs₂CO₃ (0.66 g, 2.0 mmole) in drybox. Benzophenone imine (0.32 ML, 1.9 mmole) and 30 ML of toluene wasadded under nitrogen before the mixture was heated at 90° C. for 14 h.Then it was diluted with EtOAc and brine. Organic layer was dried(Na₂SO₄) and concentrated.

[1204] Crude material was dissovled in 8 ML of MeOH and 5 ML of THFbefore the addition of NaOAc (0.56 g, 6.8 mmole) and NH₂OH-HCl (0.24 g,3.4 mmole) at RT. After 50 min, EtOAc and brine was added. Organics weredried (Na₂SO₄), concentrated and purified with silica gel (30 EtOAc/Hex)to produce crytalline 629-ys-190 (0.88 g, 1.6 mmole).

[1205] LiHMDS (1N in THF, 8.0 mmole) was added slowly to a solution of629-ys-190 (0.88 g, 1.6 mmole) in 16 ML of THF at −55 to −50° C. Themixture was stirred at −45° C. for 5 min before the addition of BOC₂O(0.38 ML, 1.8 mmole). After the mixture was stirred at −40° C. for 30min, Mel (0.60 ML, 9.6 mmole) was added. After 10 min the mixture waswarmed up to RT for 2 h. Recooled to −35° C., the solution was added 72ML of 1N NaOH and 48 ML of

[1206] EtOH. After it was heated at 45° C. for 12 h, the mixture wasdiluted with 100 ML of water and 150 ML of CH₂Cl₂. Aq. layer wasextracted twice with 50 ML of CH₂Cl₂. Organics were concentrated andpurified by silica gel chromatography (30% EtOAc/Hex) to furnishcolorless gel 629-ys-192 (0.58 g, 1.0 mmole).

[1207] The suspension of 629-ys-192 (0.40 g, 0.71 mmole), PCC (0.46 g,2.1 mmole), 4A molecular sieves (0.50 g), and celite (0.50 g) in 8 ML ofCH₂Cl₂ was stirred at RT for 2.5 h before the addition of Et₃N (0.29 ML,2.1 mmole). After 5 min, 30 ML of Et₂O was added and the mixture wasfiltered. The filtrate was concentrated and passed through a shortsilica gel plug (75% EtOAc/Hex) to provide colorless crystalline629-ys-198 (0.35 g, 0.63 mmole).

[1208] TFA (5% water, 6 ML) was added slowly to the solution of629-ys-198 (0.35 g, 0.63 mmole) in CH₂Cl₂ at −35° C. The mixture wasstirred at −20° C. for 1 h before the addition of sat. aq. NaHCO₃ (PH˜8) and CH₂Cl₂. Aq. layer was extracted twice with CH₂Cl₂. The organicswere dried (Na₂SO₄), concentrated and purified by silica gelchromatography (75% EtOAc/Hex) to afford ER-805940 (124 mg, 0.33 mmole)in 25% overall yield over 8 steps.

[1209] Synthesis of ER806201:

[1210] 1) Synthesis of Triflate:

[1211] To a solution of trihydroxy-benzoic acid (120 g) in 350 mL ofacetone, 500 mL of TFA (tri-fluoro acetic acid) was added at 40 ° C.under stirring. After 1 h at that temperature, 300 mL of TFAA(tri-fluoro acetic anhydride) was added. The mixture was heated for 3days. The mixture was distilled under house vacuum at 50 ° C. to removesolvents. The crude product was then diluted with 4 L of CH₂Cl₂, washedwith water, sat. NaHCO₃, dried and concentrated to give 85 g of semipure solid. The solid was crystallized in EtOH (1 g/2 mL) to give 20 gof pure crystal. The mother liq. was then purified by silica gel withCH₂Cl₂ to 5% MeOH/CH₂Cl₂ to give 55 g of additional product, 531-YW-184.

[1212] To a solution of 531-YW-184 (50 g, 238 mmol) in 156 mL ofpyridine, Tf₂O (100 mL, 595 mmol, 2.5 eq.) was added at 0° C. in 3 h.Then it was warmed to rt and stirred for 2 h. The reaction mixture wasdiluted with water. The mixture was filtered. The solid on the filterwas washed with water, dried under vac to give a solid, 531-YW-187 (100g).

[1213] To a mixture of ditriflate, 531-YW-187 (45.35 g), BocNH₂ (17.22g), Pd₂(dba)₃ (4.38 g) and Pt-Bu₃ (4.38 g) in 150 mL of toluene,tri-ethylamine (26.92 mL) was added. The reaction was heated under inertatmosphere at 80 ° C. for 4 h. The crude reaction mixture was cooled andfiltered through a pad of celite. The filtrates were concentrated andpurified on silica gel with Hex/EtOAc, 9:1, 4:1 to give 28.3 g ofdesired product, 531-YW-194.

[1214] 2) Synthesis of Olefin:

[1215] 792-ANDI-114A was prepared analogously to the preparation of554-RB-240 with appropriate protecting groups,i.e. the MPM ether wasreplaced with TBDPS ether.

[1216] To a solution of 792-ANDI-114A (165.9 g, 265 mmol) in 2.65 L ofHexanes, quinoline (2.65 mL) and Lindlar catalyst (28.2 g, 13. 3 mmol,0.05 eq.) were added. The mixture was degassed repeatedly under vacuumand recharged with nitrogen (3×) and hydrogen (3×). then it was set theintake of hydrogen on hydrogenator to 0.114 mol. The reaction wasmonitored by MS/1H NMR. After overnight, the suspension was filtered andrecharged with catalyst and hydrogen. After 3 days, the reaction wasfiltered through celite. The filtrates were concentrated and purified onsilica gel to give 104 g, 772RB147B as an oil.

[1217] To a solution of 772RB147B (67.4 g, 107 mmol), MPMCl (21.9 mL,161 mmol, 1.5 eq.) and a 1M solution of NaHMDS in THF (140 mL, 140 mmol,1.3 eq.) was added slowly with syringe pump in 2 h at 0° C. Afterstirred for 1.5 h at 0° C., it was quenched with sat. NH₄Cl at 0° C. andwarmed to rt. The mixture was extracted with EtOAc (3×). The extractswere washed with water, brine, dried and concentrated. The crude productwas purified on silica gel to give 772RB 162 quantitatively.

[1218] 772RB162 (119.6 g, 160 mmol) was dissolved in a mixture of 10%NaOH in methanol (3.2 L, v/v) and 3.5 mL of water. The reaction washeated at 45° C. for 48 h. After cooled, it was diluted with 9 L ofCH₂Cl₂, washed with water (2×), Sat NH₄Cl, brine, dried andconcentrated. The crude product was purified on silica gel with10%-25-35% of EtOAc/Hexanes to give the 772RB164 (78 g, 96% yield).

[1219] To a solution of (COC)₂ (25 mL, 295 mmol, 2 eq.) in CH₂Cl₂ (870mL), DMSO (41. 85 mL, 590 mmol, 4 eq.) was slowly added at −78° C. After30 min stirred at that temperature, a solution of 772RB164 (75 g, 147.4mmol) in CH₂Cl₂ (160 mL) was added in 45 min. After stirred at −78° C.for 45 min, Et₃N (82.2 mL, 590 mmol, 4 eq.) was added at thattemperature. After stirred for 30 min, it was warmed to 0° C. for 1.5 h.The reaction was quenched with 750 mL of saturated NH₄Cl and extractedwith EtOAc (3×). The extracts were dried and concentrated. The crudeproduct was re-suspended with 2.5 L of 1:1 solution of EtOAc/Hexanes,washed with water (3×), brine, dried and concentrated. The crude product772RB 169 was used directly for next step.

[1220] To a suspension of Ph₃PCH₃Br (115.8 mL, 324.3 mmol, 2.2 eq.) in amixture of THF (870 mL) and DMSO (433.6 mL), n-BuLi (184.3 mL of 1.6 Msolution, 294.8 mmol, 2 eq.) was added at 0° C. After stirred for 1 h, asolution of 772RB169 (74.7 g in 175 mL of THF, 147.4 mmol) was added at0° C. After 30 min, it was warmed to rt. After 2 h, it was quenched with1.1 L of Sat. NHCl₄ and extracted with EtOAc (3×). The extracts werewashed with water, brine and dried and concentrated. The crude productwas purified on silica gel with 5-10% EtOAc/Hexanes to give 66.5 g of772RB170 as an oil (89% yield).

[1221] 3) Coupling of Triflate and Olefin:

[1222] To a mixture of 772RB168 (2.5 g, 4.95 mmol) and triflate (2.7 g,6.4 mmol, 1.3 eq.), Pd₂(dba)₃ (1.36 g, 1.48 mmol, 0.3 eq.) was added inthe dry box. After moved out of dry box, the mixture was suspended in8.3 mL of dioxane and N-Methyl N-dicyclohexane amine (2.1 mL, 9.9 mmol,2 eq.) was added. The reaction was heated at 100° C. for 12 h withvigorous stirring. After cooled, 6 g of celite was added and dilutedwith EtOAc. The mixture was filtered through a celite plug and rinsedwith EtOAc. The filtrates were concentrated. The crude product waspurified on silica gel with 10-20% EtOAc/Hexanes to give 3 g of pure772RB172 (76% yield).

[1223] To a solution of 772RB173 (46.3 g, 58.16 mmol) in DMPU (291 mL),LiHMDS (116 mL of 1M solution in THF, 116.3 mmol, 2 eq.) was added at 0°C. After stirred for 40 min at that temperature, EtI (27.9 mL, 349 mmol,6 eq.) was added. After 5 min, it was warmed to rt. After stirred 3 h,it was quenched with 1 L of Sat. NHCl₄ at 0° C. The mixture wasextracted with MTBE/Hexanes (1:1). The extracts were washed with brine,dried and concentrated. The crude product was purified on silica gelwith 15-20% EtOAc/Hexanes to give 40 g of desired product, 77RB175 (84%yield).

[1224] To a solution of 772RB175 (48 g, 58.2 mmol) in 230 mL, a solutionof TBAF (407 mL of 1M solution, 407 mmol, 7 eq.) and imidazole.HCl (21.3g, 203.9 mmol, 3.5 eq.) was added. The reaction was heated at 60° C. for72 h. After cooled to rt, it was quenched with sat. NH₄Cl and wasextracted with ether (3×). The organic layers were washed with water,brine, dried and concentrated. The crude product was purified on silicagel with 20-30% EtOAc/Hexanes to give 31.4 g (76% yield).

[1225] To a solution of 772RB177 (20.3 g, 28.6 g) in 3 L of THF, 0.5MKHMDS solution (60 mL, 30 mmol, 1.05 eq.) was added slowly by syringepump in 120 min. After stirred for 5 min, it was quenched with 1.5 L ofsat. NH₄Cl. The mixture was extracted with ether (3×). The extracts werewashed with brine, dried and concentrated. The crude product waspurified on silica gel with 10-20%-50% EtOAc/Hexanes to give 14.2 g ofdesired product (76% yield).

[1226] To a solution of 772RB178, 179 (19 g, 29.15 mmol) in DMF (194mL), imidazole (4 g, 58.3 mmol, 2 eq.) and TBSCl (5.27 g, 35 mmol, 1.2eq.) were added. After stirred overnight, it was quenched with asaturated solution of NaHCO₃ and water. The mixture was extracted withEtOAc. The organic layer was washed with, water, brine, dried andconcentrated. The crude product was purified on silica gel column togive 22 g (99% yield) of desired product.

[1227] To a solution of 772RB181 (22 g, 28.7 mmol) in a mixture ofCH₂Cl₂ (230 mL) and H₂O (57.4 mL), DDQ (9.78 g, 43 mmol, 1.5 eq.) wasadded at 0° C. After stirred for 2 h, it was quenched with 1 L of a 1:1mixture of aq. saturated NaHCO₃ and 10% aq. Na₂S₂O₃. The mixture wasextracted with 3×1 L of ether. The extracts were washed with brine,dried and concentrated. The crude product was purified on silica gel togive 18.1 g of pure product.

[1228] To a solution of 772RB182 (18 g, 27.9 mmol) in 279 mL of CH₂Cl₂,dried 4A molecular sieves (18 g) and PCC (18 g) were added. Afterstirred for 90 min, it was quenched with Et₃N (19.45 mL). The reactionmixture was filtered through a plug of celite, the plug was rinsed with75% EtOAc in hexanes (900 mL). The filtrates were concentrated. Thecrude product was purified on silica gel column with 10-15-20%EtOAc/Hexanes to give 14.6 g (81%) pure product.

[1229] In a 2 L flask, 772RB183 (8.5 g, 13.2 mmol) was dissolved inCH₂Cl₂ (82.5 m and the mixture was cooled to 0° C. Then a solution of5%H₂0/TFA (4.1/78.1 mL) was added slowly (˜30 min) and the mixture wasstirred at 0° C. for 14.5 hrs. The reaction was monitored by TLC.Reaction mixture was diluted with CH₂Cl₂ at 0° C. Reaction was quenchedwith a solution of NaHCO₃ in water (˜1.2 eq compare to TFA). Reactionwas cooled to r.t. Extract 3× with CH₂Cl₂, dried with Na₂SO₄, Filtered,and concentrated. Chromatography on Si-Gel, 50-60-75% EtOAc/hexane gaveER-806201: 4.8 g, 93% yield.

[1230] Preparation of C5-F-Enone Series:

[1231] Preparation of ER803030:

[1232] To a magnetically stirred solution of 2-fluoro-2-phosphonoaceticacid triethylester (8 g, 33.3 mmol) in DMF (2.7 mL) at 0° C. wasintroduced sodium hydride (0.8 g, 33.3 mmol). After 1 hour of stirringat 0° C., a solution of aldehyde (3.47 g, 16.65 mmol) in THF (14 mL) wasadded. After 2.5 hours of stirring at 0° C., a saturated solution ofammonium chloride was added. The reaction mixture was diluted with waterand extracted with ethyl acetate. The crude product was purified byflash chromatography eluting with n-hexane/ethyl acetate: (20/1) toafford 496-SG-026B (3.58 g, 72% yield).

[1233] 496-SG-027B

[1234] To a magnetically stirred solution of 496-SG-026B (3.58 g, 12.1mmol) in dichloromethane (136 mL) at 0° C. was introduced DIBAL-H (1Msolution in dichloromethane, 30.2 mL, 30.2 mmol). After 0.5 hour ofstirring at 0° C., the reaction mixture was warmed-up to roomtemperature and stirred an additional 10 minutes. The reaction wascooled back to 0° C. and a saturated solution of ammonium chloride wasadded (5.4 mL). After 15 minutes of stirring, the reaction mixture wasdiluted with ether and stirred 30 minutes at room temperature. Theresulting suspension was filtered and the solid washed with ether. Thesolvent was removed by evaporation. The crude product was purified byflash chromatography eluting with hexanes/ethyl acetate: (3/1) to afford496-SG-027B (2.68 g, 87% yield).

[1235] 496-SG-028B

[1236] To a magnetically stirred solution of 496-SG-027B (2.68 g, 10.55mmol) in dichloromethane (53.2 mL) cooled to 0° C. (Ice/water, externalthermometer) was introduced DMSO (2.6 mL, 36.94 mmol) followed by P₂O₅(5.24 g, 36.94 mmol). After one hours of stirring at room temperaturethe reaction was cooled down to 0° C., and triethylamine (7.4 mL, 52.72mmol) was added. After 20 minutes of stirring at room temperature thereaction mixture was diluted with water and extracted withdichloromethane. The solvent was removed by evaporation. The crudeproduct was purified by flash chromatography eluting with hexanes/ethylacetate: (3/1) to afford 496-SG-028B (2.66 g).

[1237] 496-SG-022B

[1238] To a magnetically stirred solution of 3-butyn-1-ol (3.0 g, 42.8mmol) and imidazole (14.6 g, 214 mmol) in dichloromethane (113 mL) atroom temperature was introduced tert-butyldiphenylsilyl chloride (11.7mL). After 18 hours of stirring at room temperature the reaction wasdiluted with water and extracted with ether. The solvent was removed byevaporation. The crude product was purified by flash chromatographyeluting with hexanes/ethyl acetate: (2/1) to afford 496-SG-022B (13.7g).

[1239] 496-SG-029B

[1240] To a magnetically stirred solution of 496-SG-022B (6.5 g, 21.8mmol) in THF (208 mL) at −78° C. was introduced n-BuLi (2.5 M in hexane,8.4 mL, 21.1 mmol). After 1 hours of stirring −78° C., a solution of496-SG-028B (2.66 g, 10.55 mmol) in THF (128 mL) was added at −78° C.After 15 minutes of stirring at −78° C., the reaction was quenched byaddition of a saturated solution of ammonium chloride. The reactionmixture was diluted with water and extracted with ethyl acetate. Thesolvent was removed by evaporation. The crude product was purified byflash chromatography eluting with hexanes/ethyl acetate: (5/1) to afford496-SG-029B (4.27 g, 70%).

[1241] 496-SG-30A

[1242] A solution of 496-SG-29B (4.27 g, 7.61 mmol) and quinoline (0.033mL) in hexanes with a catalytic amount of Lindlar catalyst wasmagnetically stirred for 1 h under hydrogen atmosphere. The reactionmixture was filtered through celite, and the solvent removed byevaporation to afford 496-SG-30A (4.28 g). The crude was used in thenext step without purification.

[1243] To a magnetically stirred solution of 496-SG-030A (4.28 g7.61mmol), triethylamine (2.7 mL, 19.3 mmol) and a catalytic amount of DMAPin dichloromethane (267 mL) at room temperature, was introduced benzoylchloride (1.8 mL, 15.22 mmol). After 18 hours of stirring at roomtemperature the reaction mixture was diluted with a 0.1 M solution ofsodium hydroxide and extracted with ether. The solvent was removed byevaporation. The crude product was purified by flash chromatographyeluting with hexanes/ethyl acetate: (9/1) to afford 496-SG-031B (5.0 g,98%).

[1244] 496-SG-042B

[1245] To a magnetically stirred solution of 496-SG-031B (3.79 g, 5.68mmol) in acetone (57 mL) at room temperature, was introduced NMO (1.33g, 11.36 mmol) and water (2.9 mL). The reaction mixture was cooled downto 0° C., and a solution (0.1 M in toluene) osmium tetraoxide was added.After 18 hours of stirring at room temperature the reaction was quenchedwith sodium thiosulfate and stirred at room temperature for 20 minutes.The reaction mixture was then diluted with water and extracted withethyl acetate. The solvent was removed by evaporation. The crude productwas purified by flash chromatography eluting with hexanes/ethyl acetate:(3/1) to afford 496-SG-042B (1.5 g, 39%).

[1246] To a magnetically stirred solution of 496-SG-042B (2.16 g, 3.08mmol) and 2,2-dimethoxypropane (1.93 mL, 15.4 mmol) in a 2/1 mixtureacetone/dichloromethane (32 mL) at room temperature, was introducedcamphosufonic acid (0.8 g, 3.4 mmol). After 2 hours of stirring at roomtemperature the reaction was quenched with sodium bicarbonate and thereaction mixture was then diluted with water and extracted with ethylacetate. The solvent was removed by evaporation. The crude product waspurified by flash chromatography eluting with hexanes/ethyl acetate:(5/1) to afford 496-SG-043B (2.13 g, 93%).

[1247] 496SG-45A

[1248] To a magnetically stirred solution of 496-SG-043B (2.14 g, 2.89mmol) in THF (52 mL) at 0° C., was introduced a 1 M solution of TBAF inTHF buffered with 0.5 equivalent of imidazole hydrochloride (7.2 mL, 7.2mmol). After 1 hour of stirring at room temperature the reaction wasdiluted with water and extracted with ether. The solvent was removed byevaporation to afford 496SG-45A (1.39 g). The crude product was used inthe next step without purification.

[1249] 496-SG-046B

[1250] Using a procedure analogous to that described for the synthesisof 343-YW-281, 496-SG-045A (1.4 g, 2.79 mmol) was reacted withtriphenylphosphine (1.24 g, 4.74 mmol), DEAD (0.465 mL, 2.93 mmol) andmethyl iodide (0.225 mL, 3.63 mmol) in toluene (46.5 mL). The crudeproduct was purified by flash chromatography eluting with hexanes/ethylacetate (5/1 and then 3/1) to afford 496-SG-046B (1.46 g, 85% yield).

[1251] 496-SG-048B

[1252] Using a procedure analogous to that described for the synthesisof ER-803027 (stage 447-JCH-273B), 496-SG-046B (1.46 g, 2.38 mmol) wasreacted with intermediate 509-HD-213 (178 g, 3.00 mmol) and LiHMDS (1Msolution in THF, 3.6 mL, 3.6 mmol) in a 10/1 THF/HMPA mixture (17.3 mL)to afford after purification by flash chromatography eluting withhexanes/ethyl acetate: 496-SG-048A. Using a procedure analogous to thatdescribed for the synthesis of 447-JCH-275B, 496-SG-048A was reactedwith MCPBA (0.75 g, 2.38 mmol) and triethylamine (2 mL, 14.3 mmol). Thecrude product was purified by flash chromatography eluting withhexanes/ethyl acetate (5/1 and then 3/1) to afford 496-SG-048B (1.38 g,72% yield).

[1253] 496-SG-052B

[1254] 496-SG-48B (1.28 g, 1.58 mmol) was reacted with DDQ (0.43 g, 1.89mmol) in a 2/1-dichloromethane/water mixture (68 mL). The crude productwas purified by flash chromatography eluting with hexanes/ethyl acetate(5/1 and then 3/1) to afford 496-SG-052B (0.88 g, 81% yield).

[1255] 496-SG-053A

[1256] Using a procedure analogous to that described for the synthesisof ER-803064 (stage 509-HD-116), 496-SG-052B (0.88 g, 1.28 mmol) wasreacted with TBAF (1.0 g, 3.82 mmol) in THF (2.4 mL) to afford496-SG-053A (0.74 g). The crude product was used in the next stepwithout purification.

[1257] 496-SG-058B

[1258] 496-SG-053A (0.20 g, 0.34 mmol) was reacted withtriphenylphosphine (0.107 g, 0.408 mmol) and DEAD (0.064 mL, 0.408 mmol)in THF (90 mL). The crude product was purified on silica gel elutingwith n-hexane/ethyl acetate: (2/1) to afford 496-SG-058B (0.12 g, 62%yield).

[1259] 496-SG-057A, 496-SG-057B, 496-SG-057C

[1260] Using a procedure analogous to that described for the synthesisof ER-803064, 496-SG-058B (0.048 g, 0.084 mmol) was reacted with sodiumhydroxide (1M solution, 0.42 mL, 0.42 mmol) in a 2/1 mixture ethanol/THF(1 mL). The crude product was purified on silica gel (TLC) eluting withhexanes/ethyl acetate: (1/1) to afford 496-SG-057A (0.011 g),496-SG-057B (0.013 g), 496-SG-057C (0.01 g).

[1261] 496-SG-061A, 496-SG-061B, 496-SG-061C.

[1262] 496-SG-057A (0.01 g, 0.021 mmol), 496-SG-057B (0.012 g, 0.026mmol), 496-SG-057C (0.0095 g, 0.02 mmol) were separately reacted withDess-Martin reagent: (0.055 g, 0.129 mmol), (0.065 g, 0.154 mmol),(0.052 g, 0.122 mmol) and sodium carbonate (0.027 g), (0.032 g), (0.026g) in dichloromethane (1.5 mL), (1.8 mL), (1.4 mL). After work-up, eachreaction mixture was purified on silica gel (TLC) eluting withhexanes/ethyl acetate: (2/1) to afford respectively 496-SG-061A (0.009g), 496-SG-061B (0.006 g), and 496-SG-061C (0.011 g).

[1263] 496-SG-067A/ER-803029, 496-SG-067B/ER-803026,496-SG-067C/ER-803030.

[1264] Using a procedure analogous to that described for the synthesisof ER-803064 (final step), 496-SG-061A (0.007 g, 0.016 mmol),496-SG-061B (0.004 g, 0.009 mmol), and 496-SG-061C (0.013 g, 0.027 mmol)were separately reacted with HF 48%: (0.37 mL), (0.21 mL), (0.63 mL) inacetonitrile/dichloromethane (4/1): (1.9 mL), (1.0 mL), (3 mL). Afterwork-up, each reaction mixture was purified on silica gel (TLC) elutingwith hexanes/ethyl acetate: (2/1) to afford respectively496-SG-067A/ER-803029 (0.006 g), 496-SG-067B/ER-803026 (0.002 g), and496-SG-067C/ER-803030 (0.006 g).

[1265] ER803916

[1266] To a solution of (EtO)₂POCHFCO₂Et (5.6 mL) in a mixture of THFand DMF (100 and 25 mL), NaH (1.2 g) was added at 0° C. After stirred at0° C. for 1 h, it was cooled to −40° C. with dry ice-acetone bath. Asolution of aldehyde (3.5 g) in THF (20 mL) was added drop wise. Then,the reaction was warmed up to rt overnight. It was quenched with sat.NH₄Cl, and extracted with EtOAc (2×). The organic layers were washedwith brine, dried and concentrated under vacuum. The crude product waspurified on silica gel with 15:1, Hexanes:EtOAc to give 3.5 g of desiredproduct, 531-yw-11, which gave satisfactory ¹H NMR. The cis:trans ratioof the two isomers was 12:1 based on ¹H NMR.

[1267] To a solution of ester, 531-YW-11, (3.5 g) in ether (300 mL), asolution of DIBAL-H (15 mL) was added at 0° C. After 1 h at 0° C., itwas quenched with 4 mL of MeOH, and 15 mL of aq. Sat. Na₂SO₄. Afterstirred for 5 hr at rt, it was filtered through a pad of celite, and thepad was washed with ether (2×). The combined filtrates were concentratedto dryness to give 3.5 g of crude product, 531-YW-12.

[1268] To a solution of alcohol (5.6 g) and MPMOTCI (24 g) in 120 mL ofEt₂O, a solution of TfOH (20 mL, 0.3 mL dissolved in 25 mL of Et₂O) wasadded in 4 h. Then it was quenched with Sat. NaHCO₃ and extracted withEt₂O (2×). The organic layers were washed with brine, dried andconcentrated. The crude solid was suspended with pentanes. Theprecipitation was filtered. The filtrates were concentrated to give 15 gof crude product. It was purified on silica gel with 8:1, Hexanes/EtOActo give 12.7 g of the desired product, 531-YW-14

[1269] To a solution of (COCl)₂ (5 mL) in 250 mL of CH₂Cl₂, DMSO (10 mL)was added at −78° C. After 15 min at −78° C a solution of 531-YW-12 (3.8g) in 50 mL of CH₂Cl₂ was added at that temperature. After 30 min at−78° C., Et₃N (15 mL) was added, the reaction was warmed to 0° C. Thereaction was quenched with sat. NH₄Cl, extracted with EtOAc. The organiclayer was washed with brine, dried and concentrated. The crude productwas passed through a short silica gel pad with Hexanes/EtOAc, 8:1 togive 4.1 g of slightly impure product.

[1270] To a solution of acetylene, 531-YW-14 (3.56 g, 18.7 mmol, 1eq.)in 200 mL of THF, a solution of n-BuLi (12.9 mL, 20.58, 1.1 eq., 1.6mmol) was added at −78° C. The reaction was warmed to 0° C. for 5 min(by internal temperature). Then it was cooled back to −78° C. A solutionof aldehyde, 531-YW-12 (9.8 mmol, 0.5 eq.) in 50 mL of THF was added.The reaction was allowed to be warmed up to 0° C. in 1 h. It wasquenched with sat. NH₄Cl and purified as described before to give thedesired acetylenic alcohol, 531-YW-15.

[1271] The starting material (531-YW-15, 5.7 g, 10.2 mmol) was dissolvedin 200 mL hexanes. Quinoline (500 pL) and Lindlar catalyst (10 g) wereadded. The reaction mixture was stirred at room temperature under H₂balloon atmosphere for 1 h. Then the catalyst was filtered away.Quantitative amount of 509-HD-134 was obtained as colorless oil.

[1272] 509-HD-134 (5.7 g, 10.2 mmol) was dissolved in 100 mLdichloromethane at room temperature. Triethylamine (3.5 mL, 25.5 mmol),benzoyl chloride (2.4 mL, 20.4 mmol) and catalytic amount of DAMP wereadded, respectively. After stirring for 1 h, 0.1N sodium hydroxidesolution was added and the reaction mixture was extracted with ethylacetate. The crude product was purified on silica gel column, giving509-HD-135 as colorless oil in 77% yield.

[1273] 509-HD-135 (5.2 g, 7.64 mmol) was dissolved in acetone and water(1: 0.05) at 0° C. 4-Methylmorpholine N-oxide (1.8 g, 15.28 mmol) andsolution of osmium tetraoxide (0.1 M, 7.6 mL) in toluene were added. Thereaction mixture was warmed up to room temperature and stirred for 20 h.It was quenched with 10% sodium thiosulfate in sat. sodium bicarbonateaqueous solution, and extracted with ethyl acetate. After purificationon silica gel column, 509-HD-138 was obtained in 93% yield.

[1274] 509-HD-138 (5.1 g, 7.13 mmol) was dissolved in 80 mL ofdichloromethane. 2-Methoxypropene (1.4 mL, 14.26 mmol) and catalyticamount of pyridinium p-toluenesulfonate were added. After stirring atroom temperature for 20 min, the reaction mixture was quenched with sat.sodium bicarbonate solution and extracted with dichloromethane. Afterpurification on silica gel column, 509-HD- 139 was obtained in 90%yield.

[1275] 509-HD-139 (2.55 g, 3.38 mmol) was dissolved in the mixture of 30mL of dichloromethane and 15 mL of water.2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (844 mg, 3.72 mmol) was added.After stirred at room temperature for 1 h, the reaction mixture wasquenched with sat. sodium bicarbonate solution and extracted with ethylacetate. After purification on silica gel column, 509-HD-140 wasobtained as white foam in 98% yield.

[1276] 509-HD-140 (1.86 g, 2.93 mmol) was dissolved in 30 mL of tolueneat room temperature. Triphenylphosphine (1.31 g, 4.98 mmol) was added,followed by methyl iodide (236 μL, 3.80 mmol) and diethylazodicarboxylate (508 μL, 3.22 mmol). After stirring for 10 min, thereaction mixture was triturated with toluene. After purification onsilica gel column, 509-HD-141 was obtained as colorless oil in 96%yield.

[1277] 509-HD-141 (2.03 g, 2.73 mmol) and 509-HD-213 (2.84 g, 5.46 mmol)were dissolved in a mixture of 35 mL of THF/HMPA(10/1) at −78° C. Thesolution of LiHMDS (1N, 4.6 mL) in hexanes was added. The reactionmixture was stirred for 30 min, and then it was quenched with sat.ammonium chloride and extracted with ethyl acetate. After purificationon silica gel column, 509-HD-143 was obtained in 85% yield.

[1278] 509-HD-143 (2.85 g, 2.59 mmol) was dissolved in 50 mL ofdichloromethane at 0° C. 3-Chloroperbenzoic acid (50%, 1.8 g) was added.After stirring at 0° C. for 20 min, triethylamine (2.2 mL, 15.5 mmol)was added, then the reaction mixture was warmed up to room temperatureand stirred for 30 min. It was quenched with 10% sodium thiosulfate insat. sodium bicarbonate aqueous solution, and extracted withdichloromethane. After purification on silica gel column, 509-HD-144 wasobtained as white foam in 63% yield.

[1279] 509-HD-144 (1.33 g, 1.41 mmol) was dissolved in 10 mL of THF. TheTHF solution of TBAF buffered with imidazole.HCl (1N, 14.1 mL) wasadded. The reaction mixture was heated at 50° C. for 72 h. It wasdiluted with Et₂O and washed with H₂0. After purification on silica gelcolumn, 509-HD-150 was obtained as pale yellow foam in 95% yield.

[1280] 2-Chloro-1-methylpyridinium iodide (1.03 g, 4.02 mmol) and n-Bu₃N(958 μL, 4.02 mmol) were dissolved in 80 mL of dichloromethane andheated to reflux. The solution of 509-HD-150 (807 mg, 1.34 mmol) in 50mL of dichloromethane was added slowly. The reaction mixture was heatedfor 30 min. It was washed with 0.02N hydrochloric acid, sat. sodiumbicarbonate solution and brine, respectively. After purification onsilica gel column, 509-HD-152 was obtained in 50% yield.

[1281] 509-HD-152 (390 mg, 0.67 mmol) was dissolved in 10ml ethylalcohol. Sodium hydroxide (1N, 6.7 mL) solution was added. The reactionmixture was stirred for 1 h at room temperature. It was diluted withH₂O, extracted with EtOAc. After purification on silica gel column, 134mg of the major desired single isomer 509-HD-153C was obtained ascolorless oil.

[1282] 509-HD-153C (94.4 mg, 0.20 mmol) was dissolved in 8 mL ofdichloromethane. Molecular sieve (4A, 423 mg) and PCC (423 mg, 2.0 mmol)were added. The reaction mixture was stirred for 12 h at roomtemperature. After passing through celite, 509-HD-158 was obtained ascolorless oil in 52% yield.

[1283] 509-HD-158 (49.2 mg, 0.10 mmol) was dissolved in 1.0 mL ofdichloromethane. Then hydrofluoric acid (6N, 4 mL) was added. Thereaction mixture was stirred at room temperature for 30 min. It wasdiluted with more dichloromethane, washed with water and sat. sodiumbicarbonate solution. After purification on a plug of silica gel,ER803916 was obtained as a white solid in 81% yield.

[1284] Preparation of Intermediate for the Synthesis of ER806821:

[1285] The C14-TMS-ethyl intermediate was prepared from C4-TMS ethylselenide in analogous to the above sequences in similar yields.

[1286] The phenol was prepared using previous describe conditions forthe C14 modification series:

[1287] Preparation of ER806821:

[1288] To a solution of phenol (18.8 g, 33 mmol) in CH₂Cl₂ (300 mL) at0° C. was added Et₃N (11.5 mL, 82.5 mmol). Then, Tf₂O (8.3 mL, 49.5mmol) in CH₂Cl₂ (35 mL) was added dropwise over a period of 60 min andthe reaction was stirred for an additional 30 min. The reaction wasquenched at 0° C. with a saturated solution of NaHCO₃ (200 mL),extracted 3 times with 300 mL of CH₂Cl₂, the combined organic layerswere dried with Na₂SO₄, the solid was filtered and the solvent wasevaporated under vacuo. The crude compound was purified by flashchromatography on silica gel using 10% EtOAc/hexane as eluent to give20.1 g (28.6 mmol, 87%) of 640-RB-297 as white foam.

[1289] In a glove box, under N₂, a 200 mL round-bottom flask equippedwith a magnetic stirring bar was charged with 640-RB-297 (3.0 g, 4.27mmol), N-Methyl-N-Boc amine (784 mg, 5.98 mmol),Tris(dibenzylideneacetone)-dipalladium (196 mg, 0.21 mmol),2-di-t-butylphosphino-biphenyl (127 mg, 0.427 mmol), sodium-t-butoxide(574 mg, 5.98 mmol) and anhydrous toluene (100 mL). The flask wasremoved from the glove box and the mixture was stirred at 80° C. for 4hrs. Then the mixture was allowed to cool to room temperature, 100 mL ofa saturated solution of NH₄Cl was added and the mixture was stirred for1 hr. The mixture was extracted 3 times with 100 mL of EtOAc, theorganic layers were mixed together, dried with Na₂SO₄, the solid wasfiltered through a short plug of Si-Gel, rinsed with EtOAc and thesolvent was evaporated under vacuo. The crude compound was purified byflash chromatography on silica gel using 20% EtOAc/hexane as eluent togive 1.67 g (2.44 mmol, 57%) of 772-RB-20 as white foam.

[1290] To a solution of 772-RB-20 (1.67 g, 2.44 mmol) in EtOH (75 mL)and THF (5 mL) was added a solution of 1N NaOH (24.4 mL, 24.4 mmol) andthe resulting mixture was stirred at room temperature for 4 hrs. Then,EtOH was partially evaporated, the mixture was diluted with 500 mlEtOAc, washed with water (2×250 mL) and brine (250 mL), the organicphase was dried with Na₂SO₄, the solid was filtered and the solvent wasevaporated. The crude compound was purified by flash chromatography onsilica gel using 20-30% EtOAc/hexane as eluent to give 620 mg (1.07mmol, 44%) of 772-RB-21A and 370 mg (0.64 mmol, 26%) of 772-RB-21B(undesired stereochemistry at C8-C9) as white foams.

[1291] To a solution of 772-RB-21A (610 mg, 10.05 mmol) in CH₂Cl₂ (25mL) was added pyridine (0.425 mL, 5.25 mmol) and then Dess-Martinperiodinane (1.10 g, 2.6 mmol). The mixture was stirred at roomtemperature for 1.5 hr. It was then diluted with 100 mL of Et₂O, thesolid was removed by filtration through celite and rinsed with 100 mL ofEt₂0. The organic extract was dropped in a solution of 10% w/v Na₂S₂O₄in saturated NaHCO₃ solution and stirred for 15 min. The phases wereseparated, the organic layer was washed with 100 mL of brine, dried withNa₂SO₄, the solid was filtered and the solvent was evaporated. The crudecompound was purified by flash chromatography on silica gel using 30%EtOAc/hexane as eluent to give 560 mg (0.97 mmol, 92%) of 772-RB-22 aswhite foam. The other diastereomer 772-RB-21B was treated the same wayto give 325 mg (0.56 mmol, 91%) of 772-RB-25 as white foam.

[1292] A solution of 772-RB-22 (140 mg, 0.242 mmol) in CH₂Cl₂ was cooleddown to −35° C. in a bath of dry ice/acetone. Then a solution of 5%H₂O/TFA was added slowly. The mixture was warmed up slowly to −23° C.and stirred at this temperature for 50 min. Then the mixture was cooleddown to −35° C., neutralized with a saturated solution of NaHCO₃ andallowed to warm up to room temperature The mixture was extracted 3 timeswith CH₂Cl₂, the combined organic layers were dried with Na₂SO₄, thesolid was filtered and the solvent evaporated. This reaction wasrepeated 4 times. The crude compound was purified by flashchromatography on silica gel using 40% EtOAc/hexane as eluent to give161 mg (0.42 mmol, 42%) of ER-806821. The other diastereomer 772-RB-25was deprotected the same way and then the C8-C9 diol was isomerized bytreatment with H₂O and Silica gel in CH₂Cl₂ overnight to give 70 mg(0.178 mmol, 32% 2 steps) of ER-806821.

[1293] Alternative Synthesis of ER806821 and Synthesis of ER807563:

[1294] Synthesis of Acyclic Intermediate 20:

[1295] Preparation of Aldehyde 29:

[1296] Aldehyde 29 was prepared according to the procedure describedpreviously.

[1297] Preparation of Alkyne 30:

[1298] To a solution of 3-butyn-1-ol (8.98 g, 128 mmol, 1.0 eq.) indichloromethane (200 mL) was added triethylamine (23.5 mL, 167 mol, 1.3eq.) and 4-(dimethylamino)-pyridine (25 mg, 0.205 mmol, 0.0016 eq.). Theresulting mixture was cooled down to 0° C., added slowlytrimethyl-acetyl chloride (17.5 mL, 141 mmol, 1.1 eq.). Once exothermwas over, the reaction mixture was allowed to warm up to rt. Stirredovernight at rt. A saturated solution of NaHCO₃ was added, the biphasicmixture was stirred at rt during 1 h. Layers were separated. The aqueousone was extracted with dichloromethane. The organic layers were combinedand dried with sodium sulfate, filtered and concentrated to dryness. Thecrude oil was purified by filtration on a pad of silica gel 230-400 Meshusing 20% ethyl acetate/hexane to elute. This procedure afforded alkyne30, 17.3 g, pale yellow oil, 88% yield.

[1299] Preparation of Carbinol 31:

[1300] Zinc triflate (1.73 g, 7.31 mmol, 1.1 eq.) and (1S,2R)-(+)-N-methylephedrine (1.43 g, 7.90 mmol, 1.2 eq.) were added to a100 mL round bottom flask in a dry box. The flask was transfered to afume hood for the addition of toluene (20 mL) andN,N-diisopropylethylamine (1.39 ml, 7.94 mmol, 1.2 eq.). The resultingmixture was stirred 2 h at rt at which point alkyne 30 (1.23 g, 7.97mmol, 1.2 eq.) was added. Stirred 60 min at rt, added aldehyde 29 (1.73g, 6.64 mmol, 1.0 eq.) and stirred at rt 40 min. Quenched by theaddition of a saturated solution of ammonium chloride. Layers wereseparated, the aqueous one was extracted with Et₂O, the organic layerswere combined, washed with brine, dried with sodium sulfate, filteredand concentrated to dryness. The resulting crude oil was purified bychromatography on silica gel 230-400 Mesh using the following solventgradient: 8%, 12% and 16% ethyl acetate/hexane to elute. This procedureafforded carbinol 31, 2.21 g, colorless oil, 80% yield, 94% de.

[1301] Preparation of Alkene 32:

[1302] To a solution of carbinol 31 (2.60 g, 6.27 mmol, 1.0 eq.) inn-heptane (70 mL) was added quinoline (0.16 mL, 1.33 mmol, 0.21 eq.) andLindlar catalyst (640 mg). The resulting heterogeneous mixture wasstirred at rt during 1.5 h under a positive atmosphere of hydrogen.After that time, the reaction was found complete. It was then filteredon Celite and the filtrate was washed with a dilute HCl solution(prepared by mixing HCl 1 N (6.0 mL) with water (24 mL)) to removequinoline. Layers were separated, the organic one was washed with water(3×20 mL), dried with sodium sulfate, filtered and concentrated todryness to afford alkene 32, 2.56 g, colorless oil, 98% yield.

[1303] Preparation of Ether 33:

[1304] To a solution of alkene 32 (2.40 g, 5.76 mmol, 1.0 eq.) and thetrichloroimidate of 4-methoxybenzyl alcohol (2.05 g, 7.20 mmol., 1.25eq.) in dichloromethane (7.5 mL) was added at rt pyridiniump-toluenesulfonate (74 mg, 0.29 mmol, 0.05 eq.). The reaction mixturewas stirred at rt overnight. The reaction was found incomplete and inorder to push it to completion it required sequential additions of extrapyridinium p-toluenesulfonate (74 mg, 0.29 mmol, 0.05 eq.) andtrichloroimidate of 4-methoxy-benzyl alcohol (2.05 g, 7.20 mmol., 1.25eq.) and prolonged stirring at 40° C. The reaction was quenched byadding a 1:1 mixture of THF and water (10 mL) and stirring was continuedfor 1 h at 40° C. Dichloromethane was added, layers were separated, theorganic one was washed with NaHCO₃, dried with sodium sulfate, filteredand concentrated to dryness. The crude oil was purified bychromatography on silica gel 230-400 Mesh using the following solventgradient: 5% and 7.5% ethyl acetate/hexane to elute. This procedureafforded ether 33, 2.60 g, colorless oil, 84% yield.

[1305] Preparation of Diol 34:

[1306] To a solution of ether 33 (2.20 g, 4.10 mmol, 1.0 eq.) in THF (28mL) was added a solution of 4-methyl morpholine N-oxide (990 mg, 8.20mmol, 2.0 eq.) in water (28.0 mL). The mixture was added at rt asolution of osmium tetroxide (0.55 mL, 0.055 mmol, 0.013 eq., osmiumtetroxide 0.1 M in water). The reaction mixture was stirred at rt during19.5 h. Then it was quenched by the addition of 20 mL of a 1:1 solutionof NaHCO₃ saturated and 10% Na₂S₂O₃ in water. Stirring was continuedduring 60 min at rt. Layers were separated; the aqueous one wasextracted with ethyl acetate. The organic layers were combined, washedwith brine, dried with sodium sulfate, filtered and concentrated todryness. The resulting crude oil was purified by chromatography onsilica gel 230-400 Mesh using the following solvent gradient: 15% and40% ethyl acetate/hexane to elute. This procedure afforded diol 34, 2.06g, colorless oil, 88% yield beta/alpha ratio >=10:1.

[1307] Preparation of Acetonide 35:

[1308] To a solution of diol 34 (2.17 g, 3.80 mmol, 1.0 eq.) and2-methoxypropene (0.77 mL, 7.64 mmol, 2.0 eq.) in dichloromethane (40mL) was added at rt pyridinium p-toluenesulfonate (10 mg, 0.039 mmol,0.01 eq.). The reaction mixture was stirred at rt during 45 min andquenched by the addition of a saturated solution NaHCO₃. Dichloromethanewas added, layers were separated, the aqueous one was extracted withdichloromethane, the organic layers were combined, washed with brine,dried with sodium sulfate, filtered and concentrated to dryness. Thecrude oil was purified by chromatography on silica gel 230-400 Meshusing 10% ethyl acetate/hexane to elute. This procedure affordedacetonide 35, 1.80 g, a colorless oil, 78% yield.

[1309] Preparation of Alcohol 36:

[1310] To a solution of acetonide 35 (940 mg, 1.54 mmol, 1.0 eq.) inmethanol (15 mL) at rt was added potassium carbonate (261 mg, 1.85 mmol,1.2 eq.). The resulting mixture was stirred at 50° C. during 21 h,cooled down to rt, added n-hexane (30 mL), water (15 mL) and a saturatedsolution of NH₄Cl (30 mL). Layers were separated, the aqueous one wasextracted with ethyl ether/hexane, the organic layers were combined,dried with sodium sulfate, filtered and concentrated to dryness. Theresulting crude oil was purified by chromatography on silica gel 230-400Mesh using the following solvent gradient: 20%, 60% and 80% ethylacetate/hexane to elute. This procedure afforded alcohol 36, 787 mg, acolorless oil, 97% yield.

[1311] Preparation of Aldehyde 37:

[1312] To a solution of dimethyl sulfoxide (0.44 mL, 6.19 mmol, 4.0 eq.)in dichloromethane (5.0 mL) at −78° C. was added oxalyl chloride (1.54mL, 3.08 mmol, 2.1 eq., oxalyl chloride 2.0 M in dichloromethane). Theresulting mixture was stirred 30 min at −78° C., added alcohol 36 (787mg, 1.49 mmol, 1.0 eq.) in dichloromethane (2.0 mL to dissolve alcoholand 2×1.5 mL to rinse flask). The reaction mixture was stirred 60 min at−78° C., added triethylamine (0.87 mL, 6.22 mmol, 4.2 eq.), stirred 5min at −78° C., then warmed up to rt and stirring was continued at thattemperature during 50 min. At that point a saturated solution of NH₄Clwas added. Layers were separated and the aqueous one was extracted threetimes with dichloromethane. The organic layers were combined, dried withsodium sulfate, filtered and concentrated to dryness. The crude oil wasdissolved in a 1:1 mixture of ethyl acetate/hexane (100 mL) and washedwith water (10 mL) three times, then washed with brine, dried withsodium sulfate, filtered and concentrated to dryness. The concentratedmaterial was azeotroped with toluene and pumped to constant weight toafford aldehyde 37, 791 mg, a colorless oil, quantitative yield.

[1313] Preparation of Alkene 20:

[1314] To a suspension of methyltriphenylphosphonium bromide (549 mg,1.51 mmol, 2.2 eq.) in dimethyl sulfoxide (1.0 mL) and THF (3.6 mL) at0° C. was added n-butyllithium (0.855 mL, 1.37 mmol, 2.0 eq.,n-butyllithium 1.6 M in THF). The resulting mixture was stirred 60 minat 0° C., added aldehyde 37 (359 mg, 0.684 mmol, 1.0 eq.) in THF (2.0 mLand 3×1.0 mL for rinse). Stirred 20 min at 0° C., allowed to warm up tort and stirred 1.5 h at that temperature. Quenched by the addition of asaturated solution of NH₄Cl, layers were separated and the aqueous onewas extracted with a 1:1 mixture of ethyl ether/hexanes. The organiclayers were combined, washed with brine, dried with sodium sulfate,filtered and concentrated to dryness. The resulting crude oil waspurified by chromatography on silica gel 230-400 Mesh using thefollowing solvent gradient: 5% and 6.5% ethyl acetate/hexane to elute.This procedure afforded alkene 20, 322 mg, a colorless oil, 90% yield.

[1315] B) Synthesis of ER-806821:

[1316] Preparation of Heck-Coupling Product 22:

[1317] Alkene 20 (380 mg, 0.727 mmol, 1.0 eq.) and triflate 21 (321 mg,0.727 mmol, 1.0 eq.) were combined and addedtris-(dibenzylideneacetone)-dipalladium-(0) (33.2 mg, 0.036 mmol, 0.05eq.) under inert atmosphere (dry box). Those reagents were moved out ofthe dry box and added at rt acetonitrile (3.0 mL) and triethylamine(0.205 mL, 1.46 mmol, 2.0 eq.). The resulting mixture was stirred at 65°C. during 19 h, cooled down to rt and filtered on Celite. The filtratewas concentrated to dryness and purified by chromatography on silica gel230-400 Mesh using the following solvent gradient: 5%, 15%, 20% and 30%ethyl acetate/hexane to elute. This procedure afforded Heck-couplingproduct 22, 384 mg, white foam, 65% yield and unreacted alkene 20, 63mg, 17%.

[1318] Preparation of N-Methylaniline 23:

[1319] To a solution of aniline 22 (178 mg, 0.219 mmol, 1.0 eq.) in THF(0.34 mL) at 0° C. was added lithium hexamethyldisalazide (0.66 mL, 0.66mmol, 3.0 eq., lithium hexamethyldisalazide 1.0 M in THF). The resultingmixture was stirred at 0° C. during 60 min and added methyl iodide(0.085 mL, 1.35 mmol, 6.0 eq.). The reaction mixture was stirred 10 minat 0° C., then allowed to warm up to rt and stirred at that temperatureduring 21 h. A saturated solution of NH₄Cl and methyl-t-butyl ether wereadded. Layers were separated and the aqueous one was extracted twicewith methyl-t-butyl ether. The combined organic layers were washed withbrine, dried with sodium sulfate, filtered and concentrated to dryness.The resulting foam was purified by chromatography on silica gel 230-400Mesh using the following solvent gradient: 15% and 22% ethylacetate/hexane to elute. This procedure afforded N-methylaniline 23, 148mg, white foam, 82% yield.

[1320] Preparation of Macrocyclic Precursor 24:

[1321] To N-methylaniline 23 (119 mg, 0.143 mmol, 1.0 eq.) was added asolution of TBAF (1.0 mL, 1.0 mmol, 7.0 eq.) buffered with imidazolehydrochloride. That buffered solution of TBAF was prepared by adding 5.3g of imidazole hydrochloride to 100 mL of TBAF 1.0 M in THF and bystirring till complete dissolution. The reaction mixture was stirred at65° C. during 16 h, cooled down to rt, added a saturated solution ofNH₄Cl and methyl-t-butyl ether. The layers were separated and theaqueous one was extracted twice with methyl-t-butyl ether. The combinedorganic layers were washed with brine, dried with sodium sulfate,filtered and concentrated to dryness. The resulting foam was purified bychromatography on silica gel 230-400 Mesh using the following solventgradient: 30% and 50% ethyl acetate/hexane to elute. This procedureafforded macrocylclic precursor 24, 94.0 mg, white foam, 92% yield.

[1322] Preparation of Macrolactone 25:

[1323] To a solution of macrocyclic precursor 24 (47.0 mg, 0.0658 mmol,1.0 eq.) in THF (6.6 mL) was added potassium hexamethyldisalazide 0.5 Min toluene (0.265 mL, 0.133 mmol, 2.0 eq.). Stirred 10 min at rt,quenched with a saturated solution of NH₄Cl to bring pH to neutral.Methyl-t-butyl ether was added, layers were separated, and the aqueousone was extracted with methyl-t-butyl ether. The combined organic layerswere washed with brine, dried with sodium sulfate, filtered andconcentrated to dryness. The resulting foam was purified bychromatography on silica gel 230-400 Mesh using the following solventgradient: 15% and 25% ethyl acetate/hexane to elute. This procedureafforded macrolactone 25, 23.5 mg, white foam, 55% yield.

[1324] Preparation of Ether 26:

[1325] To a solution of macrolactone 25 (17.9 mg, 0.0272 mmol, 1.0 eq.)in dichloromethane (1.0 mL) at 0° C. was added N,N-diisopropylethylamine(0.125 ml, 0.712 mmol, 26.2 eq.) and MOMCl (0.042 mL, 0.550 mmol, 20.0eq.). The reaction mixture was allowed to warm up to rt and stirred atthat temperature during 3.75 h at which point it was quenched with asaturated solution of NH₄Cl. Methyl-t-butyl ether was added, layers wereseparated, and the aqueous one was extracted twice with methyl-t-butylether. The combined organic layers were washed with brine, dried withsodium sulfate, filtered and concentrated to dryness. The resulting foamwas purified by chromatography on silica gel 230-400 Mesh using thefollowing solvent gradient: 30% and 45% ethyl acetate/hexane to elute.This procedure afforded ether 26, 18.2 mg, white foam, 96% yield.

[1326] Preparation of Allylic Alcohol 27:

[1327] To a solution of ether 26 (18.2 mg, 0.026 mmol, 1.0 eq.) indichloromethane (0.60 mL) at rt was added water (0.12 mL) and DDQ (12.0mg, 0.0517 mmol, 2.0 eq.). The resulting mixture was stirred at rtduring 3.5 h at which point it was cooled down to 0° C. and quenchedwith a 1:1 solution of NaHCO₃ saturated and 10% Na₂S₂O₃ in water.Methyl-t-butyl ether was added, layers were separated, and the aqueousone was extracted twice with methyl-t-butyl ether. The combined organiclayers were washed with brine, dried with sodium sulfate, filtered andconcentrated to dryness. The resulting foam was purified by HPTLCprep-plates using 35% ethyl acetate/hexane to elute. This procedureafforded alcohol 27, 12.6 mg, white foam, 84% yield.

[1328] Note: Alcohol 27 is an intermediate of the first-generationsynthesis of ER-806821, its conversion into ER-806821 can be achievedusing the Dess-Martin periodinane and the TFA procedures.

[1329] C) Synthesis of ER-807563

[1330] Preparation of N-ethylaniline 39:

[1331] To a solution of aniline 22 (386 mg, 0.474 mmol, 1.0 eq.) in DMPU(1.6 mL) at 0° C. was added lithium hexamethyldisalazide (0.950 mL,0.950 mmol, 2.0 eq., lithium hexamethyldisalazide 1.0 M in THF). Theresulting mixture was stirred at 0° C. during 30 min and added ethyliodide (0.230 mL, 2.88 mmol, 6.1 eq.). The reaction mixture was stirred10 min at 0° C. then allowed to warm up to rt and stirred at thattemperature during 1 h. A saturated solution of NH₄Cl, methyl-t-butylether and n-hexanes were added. Layers were separated and the aqueousone was extracted twice with methyl-t-butyl ether/hexane (1:1 mixture).The combined organic layers were washed with brine/water (1:1 mixture),then with brine, dried with sodium sulfate, filtered and concentrated todryness. The resulting foam was purified by chromography on silica gel230-400 Mesh using the following solvent gradient: 15% and 20% ethylacetate/hexane to elute. This procedure afforded N-ethyl-aniline 39, 349mg, white foam, 87% yield.

[1332] Preparation of Macrocyclic Precursor 40:

[1333] N-ethylaniline 39 (817 mg, 0.970 mmol, 1.0 eq.) was added asolution of TBAF (6.8 mL, 6.8 mmol, 7.0 eq.) previously buffered withimidazole hydrochloride. That buffered solution of TBAF was prepared byadding 5.3 g of imidazole hydrochloride to 100 mL of TBAF 1.0 M in THFand by stirring till complete dissolution. The reaction mixture wasstirred at 65° C. during 16 h, cooled down to rt, added a saturatedsolution of NH₄Cl, water and methyl-t-butyl ether. The layers wereseparated and the aqueous one was extracted twice with methyl-t-butylether. The combined organic layers were washed with brine/water (1:1mixture), then washed with brine, dried with sodium sulfate, filteredand concentrated to dryness. The resulting foam was purified bychromatography on silica gel 230-400 Mesh using the following solventgradient: 20%, 30% and 45% ethyl acetate/hexane to elute. This procedureafforded macrocylclic precursor 40, 635 mg, white foam, 90% yield.

[1334] Preparation of Macrolactone 41:

[1335] To a solution of macrocyclic precursor 40 (303 mg, 0.416 mmol,1.0 eq.) in THF (40 mL) was added potassium hexamethyldisalazide (1.70mL, 0.85 mmol, 2.0 eq., potassium hexamethyldisalazide 0.5 M in toluene)over 3 min. The reaction mixture was stirred 10 min at rt and quenchedwith a saturated solution of NH₄Cl. Methyl-t-butyl ether and water wereadded, layers were separated, and the aqueous one was extracted withmethyl-t-butyl ether. The combined organic layers were washed with asaturated solution of NH₄Cl, then washed with brine, dried with sodiumsulfate, filtered and concentrated to dryness. The resulting foam waspurified by chromatography on silica gel 230-400 Mesh using thefollowing solvent gradient: 15% and 18% ethyl acetate/hexane to elute.This procedure afforded macrolactone 41, 134 mg, white foam, 48% yield.

[1336] Preparation of Allylic Alcohol 42:

[1337] To a solution of macrolactone 41 (146.5 mg, 0.219 mmol, 1.0 eq.)in dichloromethane (0.50 mL) was added water (0.50 mL). The resultingmixture was cooled down to 0° C. and was added a saturated solution ofDDQ in dichloromethane (3.0 mL, 0.265 mmol, 1.2 eq., this saturatedsolution of DDQ in dichloromethane contained at rt 20.5 mg of DDQ 98%pure/mL of dichloromethane). The biphasic reaction mixture was stirred 5min at 0° C., then 6 h at rt. At that point, it was cooled down to 0°C., added dichloromethane and quenched with a 1:1 solution of NaHCO₃saturated and 10% Na₂S₂O₃ in water. Stirring was continued for 15 min.Layers were separated and the aqueous one was extracted withdichloromethane. The combined organic layers were washed with a 1:1solution of NaHCO₃ saturated and 10% Na₂S₂O₃ in water, then washed withbrine, dried with sodium sulfate, filtered and concentrated to dryness.The resulting foam was purified by chromatography on silica gel 230-400Mesh using the following solvent gradient: dichloromethane, 3%, 5% and7.5% methyl-t-butyl ether/dichloromethane to elute. This procedureafforded allylic alcohol 42, 102 mg, white foam, 85% yield.

[1338] Preparation of Enone 43:

[1339] To a solution of allylic alcohol 42 (20.5 mg, 0.0372 mmol, 1.0eq.) in n-heptane (1.0 mL) at rt was added MnO₂ (16 mg, 0.184 mmol, 5eq.). The resulting heterogeneous mixture was stirred at 65° C. during12 h. This process gave only a small amount of desired material and inorder to push the reaction to completion, fresh MnO₂ (16 mg) was addedon a daily basis until the reaction went to completion after one week ofstirring at 65° C. Upon completion, ethyl acetate was added and thesolids were filtered off on a short silica gel column (230-400 Mesh)using ethyl acetate as solvent. The filtrate was concentrated down todryness and purified by chromatography on silica gel 230-400 Mesh using10% acetone/toluene as solvent. This procedure afforded enone 43, 7.0mg, white foam, 35% yield (not optimized).

[1340] Preparation of ER-807563:

[1341] Enone 43 is treated according to the same TFA protocol as for thepreparation of ER-805940.

[1342] Preparation of NF2561

[1343] To a solution of 3 (10.0 g, 44.2 mmol) in DME (500 mL), pyridine(5.36 mL, 66.3 mmol, 1.5 eq.) followed by nitronium tetrafluoroborate(8.8 g, 66.3 mmol, 1.5 eq.) were added at −60° C. in dry ice-acetonebath and stirred for 1 h at −50° C. Then, pyridine (5.36 mL, 66.3 mmol,1.5 eq.) followed by nitronium tetrafluoroborate (8.8 g, 66.3 mmol, 1.5eq.) were added again at −60° C. in dry ice-acetone bath and stirred for1 h at −50° C. One more time, pyridine (5.36 mL, 66.3 mmol, 1.5 eq.)followed by nitronium tetrafluoroborate (8.8 g, 66.3 mmol, 1.5 eq.) wereadded at −60 C in dry ice-acetone bath and stirred for 1 h at −50° C.The reaction mixture was quenched at −58° C. with sat. NH₄Cl (1 L) andextracted with EtOAc (2×1 L). The organic layers were washed with waterand brine, dried over Na₂SO₄ and concentrated in vacuo. The crudeproduct was purified on silica gel column (Merck 230-400mesh,) withhexanes/EtOAc, 3:1 to give 9.59 g (35.4 mmol, 80%) of desired product asa yellow crystal.

[1344] To a mixture of 4 (10.55 g, 38.9 mmol) and pyridine (15.7 mL, 194mmol, 5 eq.) in CH₂Cl₂ (100 mL), Tf₂O (9.8 mL, 58.3 mmol, 1.5 eq.) wasgradually added at 0° C. during 30 min and the reaction mixture wasstirred for 45 min. The reaction mixture was quenched with sat.NH₄Cl(200 mL) and extracted with EtOAc (2×500 mL). The organic layers werewashed with water and brine, dried over Na₂SO₄ and concentrated invacuo. The crude product was purified on silica gel column (Merck230-400mesh) with hexanes/EtOAc, 4:1 to give 13.9 g (34.5 mmol, 89%) ofdesired product as a yellow crystal.

[1345] A mixture of 5 (13.9 g, 34.5 mmol) and N-methylbenzylamine (22.7mL, 175.9 mmol, 5.1 eq.) in THF (175 mL) was refluxed overnight. Thereaction mixture was diluted with EtOAc (750 mL) and washed with water,10% KHSO₄, water and brine, dried over Na₂SO₄ and concentrated in vacuo.The crude product was purified on silica gel column (Merck 230-400mesh)with hexanes/EtOAc, 20:1, 9:1 to give 9.7 g (25.9 mmol, 75%) of desiredproduct as a colorless oil.

[1346] A mixture of 6 (9.2 g, 24.6 mmol) and 20% Pd(OH)₂/C (1.75 g) inEtOH (200 mL) was hydrogenated under 1 atm of hydrogen for 4 hrs. Thecatalyst was filtered off and the filtrate was concentrated in vacuo.The crude diamine was obtained as yellow oil. It was used withoutpurification.

[1347] A mixture of the crude diamine, triethyl orthoformate (13.1 mL,78.7 mmol, 3.2 eq.) and montmorillonite KSF (2.7 g) in EtOH (200 mL) wasrefluxed for 100 min. The insoluble material was filtered off and thefiltrate was concentrated in vacuo. The crude product was purified onsilica gel column (Merck 230-400mesh, 350 g) with hexanes/EtOAc, 1:1,1:2 and EtOAc to give 84% (2 steps) of desired product as a yellow oil.

[1348] To a solution of 7 (5.75 g, 21.8 mmol) in CH₂Cl₂ (125 mL), TFA(16.8 mL, 218 mmol,

[1349] 10 eq.) was added at 0° C. Then, the reaction mixture was warmedto room temperature and stirred for 1.5 hrs. The reaction mixture waspoured into ice water and neutralized with NaHCO₃ and extracted withEtOAc (800 mL+2×125 mL). The organic layers were washed with water andbrine, dried over Na₂SO₄. Na₂SO₄ was filtered and washed with CH₂Cl₂ andthe filtrate was concentrated in vacuo. The crude product was obtainedas white crystal and use without purification for the next step.

[1350] To a mixture of 60% NaH in mineral oil (2.3 g, 56.6 mmol, 2.6eq.) and DMF (40 mL), a solution of the crude phenol 8 in DMF (60 mL)was gradually added at 0° C. and stirred for 1 hr. Then, TBDPSCl (9.6mL, 37.0 mmol, 1.7 eq.) was added and the reaction mixture was warmed toroom temperature and stirred for 1.5 hr. The reaction mixture was pouredslowly in sat.NH₄Cl at 0° C. and extracted with EtOAc (3×150 mL). Theorganic layers were washed with water (×2) and brine, dried over Na₂SO₄and concentrated in vacuo. The crude product was purified on silica gelcolumn (Merck 230-400mesh) with hexane/EtOAc, 3:1, 1:1, 2:3 to give 8.89g (19.4 mmol, 89% 2 steps) of desired product as a colorless oil.

[1351] A mixture of 9 (8.39 g, 18.3 mmol), NBS (3.6 g, 20.1 mmol, 1.1eq.) and AIBN (1.2 g, 7.3 mmol, 0.4 eq.) in CCl₄ (250 mL) was heatedslowly to 50° C. and stirred for 4 hrs. The reaction mixture was cooledto room temperature and insoluble material was filtered off with celite,rinsed with CCl₄ and the filtrate was concentrated in vacuo. The crudebromide was used without purification. To a mixture of Cs₂CO₃ (9.2 g,28.4 mmol, 1.55 eq.) and DMF (90 mL), PhSH (2.9 mL, 28.4 mmol, 1.55 eq.)was added at room temperature and the reaction mixture was stirred for50 min. Then, a solution of the crude bromide in DMF (210 mL) was addedand the reaction mixture was stirred for 1 hr. The reaction mixture wasquenched with sat.NH₄Cl (300 mL) and extracted with EtOAc (3×300 mL).The organic layers were washed with water (×2) and brine, dried overNa₂SO₄ and concentrated in vacuo. The crude product was purified onsilica gel column (Merck 230-400mesh, 550 g) with hexanes/EtOAc, 2:1,1:1, 1:3 to give 8.52 g (15.0 mmol, 82% 2 steps) of desired product.

[1352] To a solution of 10 (8.47 g, 14.9 mmol) in THF (200 mL), TBAF(1.0Msolution in THF, 22.4 mL, 1.5 eq.) was added at 0° C. and thereaction mixture was warmed to room temperature and stirred for 75 min.The reaction mixture was quenched with sat. NH₄Cl (150 mL) and extractedwith EtOAc (3×200 mL). The organic layer was washed with water andbrine, dried over Na₂SO₄ and concentrated in vacuo. The crude productwas purified on silica gel column (Merck 230-400mesh, 550 g) withhexanes/EtOAc, 2:1, 1:1, 1:2 to give 4.7 g (14.3 mmol, 96%) of desiredproduct.

[1353] To a solution of phenol 11 (4.41 g, 13.4 mmol) in DMF (100 mL)were added DBU (3.0 mL, 20.1 mmol, 1.5 eq.) and MOMCl (1.5 mL, 20.1mmol, 1.5 eq.) at rt. and the reaction mixture was stirred for 1.5 hr.Then, DBU (1.5 mL, 10.1 mmol, 0.75 eq.) and MOMCl (0.75 ml, 10.1 mmol,0.75 eq.) were added and the reaction mixture was stirred for 1 hr.

[1354] The reaction mixture was quenched with sat. NH₄Cl (100 mL) andextracted with EtOAc (3×150 mL). The organic layers were washed withwater and brine, dried over Na₂SO₄ and concentrated in vacuo. The crudeproduct was purified on silica gel column (Merck 230-400mesh) withhexanes/EtOAc, 1:1, and 1:3 to give 87% of desired product

[1355] A mixture of 12 (4.5 g, 12.1 mmol), KOH 2M aq sol. (30 mL, 60.4mmol, 5 eq.) and DMSO (100 mL) was stirred at 80° C. for 1.5 hrs. Thereaction mixture was cooled and adjusted to pH5 with 1N HCl (Be careful,MOM group can be easily hydrolyzed if pH too acidic). The resultedcrystal (desired product) was filtered and washed water. Total 3.22 g ofcrude product was obtained.

[1356] 11) Esterification

[1357] To a solution of PPh₃ (2.3 eq) in THF (30 mL), DEAD (2.6 eq.) wasadded at 0° C. and the reaction mixture was stirred for 30 min. Then, amixture of 13 (1 eq) and TMS ethanol (1.5 eq.) in THF (50 mL) was addedand the reaction mixture was warmed to room temperature and stirred forovernight. Furthermore, PPh₃ (2.3 eq.) and DEAD (2.6 eq.) was added andstirred for 10 min. Then, TMS ethanol (1.5 eq.) was added and stirredfor 20 min. The reaction mixture was concentrated in vacuo. Et₂O wasadded to the residue and stirred. The resulted solid (Ph₃P(O)) wasfiltered off and the filtrate was concentrated in vacuo. The crudeproduct was purified on silica gel column (Merck 230-400 mesh) withCH₂Cl₂/MeOH, 200:1, 100:1 to give a mixture of desired product and1,2-dicarbethoxyhydrazine (not separated). This mixture was purified onsilica gel column (Merck 230-400 mesh) with hexanes/EtOAc, 2:1, 1:1, and2:3 to give the desired product as white crystal.

[1358] To a solution of 14 (730 mg, 1.59 mmol, 1.5 eq.) and iodide 15(639 mg, 1.06 mmol, 1 eq.) in dry THF (25 mL)-HMPA (2.5 mL), LiHMDS(1.0M solution in THF, 2.65 mL, 2.5 eq.) was added over a period of 25min at −50° C. and the reaction mixture was for 30 min. Then, LiHMDS(1.0M solution in THF, 1.3 mL, 1.25 eq.) was added at −50° C. and thereaction was stirred for 1 hr. The reaction mixture was quenched withsat. NH₄Cl (50 mL) and extracted with EtOAc (3×50 mL). The organic layerwas washed with water and brine, dried over Na₂SO₄ and concentrated invacuo. The crude product was purified on silica gel column (Merck230-400 mesh) with hexanes/EtOAc, 4:1, 1:1, 1:3 to give 440 mg (0.471mmol, 44%) of desired product as a colorless amorphous.

[1359] To a solution of 16 (980 mg, 1.22 mmol) in CH₂Cl₂ (45 mL), mCPBA(0.5 eq calculated if mCPBA was 100%, 91 mg) was added at 0° C. andstirred for 25 min, then mCPBA (91 mg) was added and stirred for 25 min,mCPBA (45 mg) was added and stirred for 25 min, mCPBA (35 mg) was addedand stirred for 20 min. The reaction mixture was quenched with sat.Na₂S₂O₃ (80 mL) and extracted with EtOAc (400ml+2×80 mL). The organiclayer was washed with sat. NaHCO₃ (×2) and brine, dried over Na₂SO₄ andconcentrated in vacuo to give 1.02 g of crude sulfoxide as colorlessamorphous.

[1360] A solution of crude sulfoxide and Et₃N (10 drops) in toluene (50mL) was refluxed for 1 hr.The reaction mixture was concentrated in vacuoand the crude product was purified on silica gel column (Merck 230-400mesh) with hexanes/EtOAc, 3:1, 1:1, 1:3 to give 980 mg (1.19 mmol, 98% 2steps) of desired product as colorless amorphous.

[1361] To a solution of 18 (924 mg, 1.12 mmol) in THF (25 mL),Imidazole,HCl (293 mg, 2.8 mmol, 2.5 eq) and TBAF (1.0Msolution in THF,5.6 ml, 5 eq) were added and the mixture was heated to 50° C. After 1 hrand 3 hrs, Imidazole HCl (293 mg, 2.8 mmol, 2.5 eq.) and TBAF(1.0Msolution in THF, 5.6 mL, 5 eq.) were added. After 24 hrs, TBAF(1.0Msolution in THF, 11.2 mL, 10 eq) was added and the reaction mixturewas stirred for 84 hrs at 50° C. The reaction mixture was quenched withsat. NH₄Cl (40 mL) and saturated with NaCl (10 mL) and extracted withEtOAc (5×100 mL). The organic layers were dried over Na₂SO₄ andconcentrated in vacuo. The crude product was purified on silica gelcolumn (Merck 230-400 mesh) with EtOAc, EtOAc/MeOH, 99:1, 95:5 to give740 mg of desired product (not pure) as a colorless amorphous.

[1362] To a refluxed mixture of 2-chloro-1-methylpyridinium iodide (4.21mmol, 4.0 eq.) and n-Bu₃N (1.0 mL, 4.21 mmol, 4 eq.) in CH₂Cl₂ (50 mL),a solution of 19 (640 mg, 1.05 mmol) in CH₂Cl₂ (25 mL) was addeddrop-wise with syringe pump during 1 hr and then the reaction mixturewas stirred for 30 min. The reaction mixture was concentrated in vacuoand the residue was diluted with EtOAc (750 mL), washed with water,sat.NH₄Cl, water and brine, dried over Na₂SO₄, filtered and concentratedin vacuo. The crude product was purified on silica gel column (Merck230-400 mesh) with hexanes/EtOAc, 1:1, 1:5 CH₂Cl₂/MeOH, 95:5 to give 560mg of desired product (not pure) as colorless amorphous.

[1363] The hydrolysis was carried out as it described before in thesynthesis of ER803064.

[1364] To a solution of 21 (260 mg, 0.596 mmol) in CH₂Cl₂ (45 mL),powdered Molecular Sieves 4A (640 mg) and PCC (642 mg, 2.98 mmol, 5 eq.)were added at room temperature and the reaction mixture was stirred for1 hrs. The reaction mixture was filtered through Celite and the filtratewas concentrated in vacuo. The crude product was purified on silica gelcolumn (Merck 230-400 mesh) with CH₂Cl₂/MeOH, 99:1, 95:5 to give 185 mgof desired product (including pyridinium impurity).

[1365] To a solution of 22 (185 mg, 0.382 mmol) in CH₂Cl₂ (10 mL), TFA(0.88 mL, 11.5 mmol, 30 eq.) was added at 0° C. and the reaction mixturewas warmed to room temperature and stirred for 1.5 hrs. The reactionmixture was concentrated in vacuo. The crude product was purified onsilica gel column (Merck 230-400 mesh) with CH₂Cl_(2/)MeOH, 98:2, 95:5,92:8 to give 120 mg (0.300 mmol, 79%, 50% 2 steps) of desired product,NF2561. The pure product was lyophilized with Water/MeCN, 1:1 to give awhite foam.

[1366] ER805911 and ER805977:

[1367] These analogs were prepared using appropriate alternativereagents using the above synthesis. The modified steps are describedbelow:

[1368] A mixture of the crude diamine (24.5 mmol), triethylacetylformate (13.5 mL, 3 eq.) and montmorillonite KSF (2.5 g) in EtOH(150 mL) was refluxed for 2 h. The insoluble material was filtered offand the filtrate was concentrated in vacuo. The crude product waspurified on silica gel column (Merck 230-400 mesh, 350 g) withhexane/EtOAc, 1:1, 1:2 and EtOAc to give 81% (2 steps) of desiredproduct as yellow oil. The product was carried forward in an analogousmanner as described for NF2561 to give ER805911.

[1369] A mixture of triflate (9.3 g, 23.1 mmol) and N-ethylbenzylamine(15.2 mL, 117.8 mmol, 5.1 eq.) in THF (175 mL) was refluxed 26 h. Thereaction mixture was diluted with EtOAc (750 mL) and washed with water,10% KHSO₄, water and brine, dried over Na₂SO₄ and concentrated in vacuo.The crude product was purified on silica gel column (Merck 230-400 mesh)with hexane/EtOAc, 20:1, 9:1 to give 6.25 g (16.1 mmol, 70%) of desiredproduct as a colorless oil. It was carried forwad similarly as describedfor NF2561 to give ER805977.

[1370] Preparation of C10 Analogs, ER804747:

[1371] The suspension of selenium dioxide (677 mg) and 531-YW-4 (1.96 g)in dioxane (30 mL) was kept at 70° C. for 10 hours. The mixture wasconcentrated and purified with flush chromatograph (hexanes/acetate 5/1to 2/1) to give 593-YJ-22-1 (396 mg) and 593-YJ-22-2 (683 mg).

[1372] Sodium hydride (60%, 59 mg) was added to the solution of593-YJ-22-2 (788 mg) in DMF (15 mL), followed by the addition ofmethoxymethyl chloride (160 mg) at room temperature. The mixture waskept stirring at room temperature overnight and quenched with aqueousammonium chloride. The aqueous phase was extracted with ether and thecombined organic phase was concentrated. The residue was purified byflush chromatograph (hexane/acetate 4/1) to yield 593-YJ-25 (431 mg).

[1373] The solution of 593-YJ-25 (431 mg) and sodium hydroxide (1.0 N,1.0 mL) in ethanol (5 mL) was kept stirring overnight at roomtemperature. The mixture was concentrated and diluted with aqueousammonium chloride. The aqueous phase was extracted with ether and thecombined organic phase was concentrated. The residue was purified by TLC(hexanes/acetate 2/1) to yield 593-YJ-29 (184 mg).

[1374] Dess-Martin periodinane was added to 593-YJ-29 (184 mg) inmethylene chloride (10 mL) at room temperature. The mixture was dilutedwith ether in 2 hours and filtrated through Celite. The filtrate wasconcentrated and the residue was purified by TLC (hexanes/acetate 2/1)to give 593-YJ-31 (138 mg).

[1375] n-Butyllithium (2.5 M, 0.56 mL) was added to 554-RB-228 (450 mg)in THF (5 mL) at −78° C. After one hour 593-YJ-31 (138 mg) was added.The reaction was kept at 0° C. for one hour and warmed to roomtemperature before it was quenched with aqueous ammonium chloride. Theaqueous phase was extracted with ether and the combined organic phasewas dried over sodium sulfate. The solvent was stripped off and theresidue was purified with TLC (hexane/acetate 3/1) to 593-YJ-32 (168 mg,75%).

[1376] The suspension of Rieke zink and 593-YJ-32 (168 mg) in methanol(10 mL) and water (1.0 mL) was kept at 70° C. for 4 hours. The mixturewas filtrated through Celite and dried over sodium sulfate. The organicswas concentrated and further dried azeotropically to give 593-YJ-33 (200mg). The crude 593-YJ-33 was subjected to triethylamine (2.7 mL) andbenzoyl chloride (1.1 mL) and purified with TLC (hexanes/acetate 4/1) togive 593-YJ-35 (358 mg).

[1377] The solution of 593-YJ-35 (358 mg) and imidazole hydrochloritebuffered TBAF (1.0 M, 0.96 mL) in THF (10 mL) was kept stirringovernight at 50° C., and then diluted with water. The aqueous phase wasextracted with ether and concentrated. The residue was purified with TLC(methylene chloride/methanol, 10/1) to give 593-YJ-36 (41 mg). 593-YJ-36(41 mg) was added to the reflux of 2-chloro-1-methylpyridium iodide (52mg) and tributylamine (43 mg) in methylene chloride (20 ml). After 2hours reflux the mixture was stirred overnight. The mixture was dilutedwith ether and washed with HCl (1.0 N) and water. The residue waspurified with TLC (hexane/acetate 1/1) to give 593-YJ-39-1 (4.3 mg).

[1378] The solution of 593-YJ-39 (4.3 mg) and sodium hydroxide (1.0 N,1.0 mL) in ethanol (5 mL) was kept stirring overnight at roomtemperature. The mixture was concentrated and diluted with aqueousammonium chloride. The aqueous phase was extracted with ether and thecombined organic phase was concentrated. The residue was purified by TLC(hexanes/acetate 2/1) to yield 593-YJ-57 (4.0 mg).

[1379] The suspension of MnO₂ (90%, 15 mg) and 593-YJ-57 (4.0 mg) inmethylene chloride (2 mL) was kept stirring overnight. The mixture wasfiltrated through Celite and concentrated. The residue was purified withTLC (hexanes/acetate 2/3) to give 593-YJ-58 (2.0 mg).

[1380] Hydrofluoric acid (49%, 0.6 mL) was added to 593-YJ-58 (2.0 mg)in acetonitrile (1.5 mL) and stirred for 20 minutes. The mixture wasdiluted with water and extracted with methylene chloride. The organicphase was concentrated and purified with a short silica gel pad toproduce 593-YJ-59 (0.5 mg, ER-804747).

[1381] Preparation of C15-Methoxy-Analog, NF1872

[1382] To a solution of 3,4,5-trimethoxytoluene (5.47 g, 30 mmol) in DME(80 mL), CuBr₂ (6.8 g, 30.45 mmol) was gradually added and the mixturewas stirred at room temperature for 30 min. Then, CuBr₂ (8.4 g, 37.61mmol) was added in several times. The mixture was stirred for 12 hrs.The insoluble material was filtered and the filtrate was concentrated.The crude product was purified on silica gel column with hexane/EtOAc,4:1 to give 7.5 g of NY-60.

[1383] NY-60 (7.39 g, 28.3 mmol) was dissolved in Et₂O (300 mL) andcooled to −78° C., under nitrogen. Then, n-BuLi (1.6M/hexane, 21 mL,33.6 mmol) was slowly added and the reaction was stirred at −78° C. for40 min. Dry ice was added to the solution, then the solution was allowedto warm to rt and was stirred for 15 min. The mixture was quenched withwater, acidified with 2N HCl, extracted with EtOAc (×2). The organiclayers were washed with water, brine and dried over MgSO₄, filtered andconcentrated to give 6.38 g of NY-61.

[1384] To a solution of NY-61 (6.37 g, 28.16 mmol) in DMF (200 mL),Cs₂CO₃ (9.17 g, 28.14 mmol) was added and stirred at room temperaturefor 10 min. Then, MeI (10 mL, 160.6 mmol) was added and the reactionmixture was stirred for 12 hrs. The reaction mixture was poured intoice-cooled sat. NH₄Cl and extracted with EtOAc (×3). The organic layerswere washed with water (×3), brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified on silica gel column withhexane/EtOAc, 4:1 to give 6.07 g of NY-62.

[1385] NY-62 (6.07 g, 25.26 mmol) was dissolved in CH2Cl2 (80 mL) andcooled to

[1386] −78° C., under nitrogen. Then, BCl₃ (1M/CH₂Cl₂, 26 mL, 26 mmol)was slowly added and the reaction was stirred at −78° C. for 1 hr. Thesolution was stirred at 0° C. for 15 min, at room temperature for 1 hr.To the mixture which was recooled to −78° C., additional BCl₃(1M/CH₂Cl₂, 52 mL, 52 mmol) was slowly added and the solution wasallowed to warm to room temperature and was stirred for 15 hrs. Thereaction mixture was poured into ice-water and extracted with EtOAc(×2). The organic layers were washed with water (×2), brine, dried overMgSO₄, filtered and concentrated. The crude product was purified byrecrystallization from hexane/EtOAc to give 4.1 g of NY-63.

[1387] To a mixture of NY-63 (4.09 g, 19.27 mmol), MeOH (1.77 mL),^(i)Pr₂NEt (3.7 mL, 21.24 mmol) and MeCN (80 mL),trimethylsilyldiazomethane (2M/hexane, 10.6 mL, 21.2 mmol) graduallyadded at 30° C. The reaction mixture was allowed to warm to roomtemperature and stirred for 24 hrs. The reaction mixture was quenchedwith water and diluted with EtOAc. The organic layer was washed with 10%citric acid and the aqueous layer was reextracted with EtOAc. Theorganic layers were washed with water, brine, dried over Na₂SO₄,filtered and concentrated. The crude product was purified on silica gelcolumn with hexane/EtOAc, 20:1, 15:1, 10:1 to give 2.29 g of mixture ofNY-64 and NY-65.

[1388] Using same procedure for NY-07, NY-65 was converted to NY-66(1.73 g).

[1389] (NY-64 was separated by column chromatography)

[1390] Using same procedure for 10, NY-66 (1.72 g, 6.41 mmol) wasconverted to NY-67 (2.07 g).

[1391] Using same procedure for NY-09, NY-67 (2.06 g, 5.47 mmol) wasconverted to NY-68 (1.54 g).

[1392] Using same procedure for 509-HD-209, NY-68 (1.48 g, 4.43 mmol)was converted to NY-69 (1.62 g).

[1393] Using same procedure for 509-HD-212, NY-69 (1.62 g, 4.28 mmol)was converted to NY-70 (1.53 g).

[1394] Using same procedure for 509-HD-213, NY-70 (1.51 g, 4.14 mmol)was converted to NY-71 (1.08 g).

[1395] Using same procedure for 16, NY-27 (51 1mg, 0.707 mmol) wasconverted to NY-72 (971 mg).

[1396] Using same procedure for 18, NY-72 (971 mg, 0.707 mmol) wasconverted to NY-33 (521 mg).

[1397] Using same procedure for 509-HD-116, NY-73 (521 mg, 0.549 mmol)was converted to NY-74 (444 mg). NY-74 was used without purification forthe next step.

[1398] Using same procedure for 509-HD-118, NY-74 (444 mg, 0.549 mmol)was

[1399] converted to NY-75 (86 mg) and NY-76 (114 mg).

[1400] Using same procedure for TM-13, NY-75 (45 mg, 0.094 mmol) wasconverted to NY-77 (19.2 mg).

[1401] Using same procedure for NF-0675, NY-77 (18 mg, 0.038 mmol) wasconverted to NF-1872 (12.6 mg).

[1402] Preparation of C16 Analogs: NF0934, NF1418 and NF1419

[1403] Synthetic Procedure for NF-0934

[1404] Using same procedure for 554-RB-238, 531-yw-2-2 (5 g, 19.21 mmol)was converted to NY-20 (5.08 g). NY-20 was used without purification forthe next step.

[1405] To a suspension of lithium acetylide-ethylenediamine complex(24.92 g, 0.271 mol) in DMSO (250 mL), (S)-propylene oxide (14.3 g,0.246 mol) was slowly added at 0° C. Then, the mixture was warmed toroom temperature and stirred for 24 hrs. The mixture was poured intoice-water and extracted with Et₂O (×4). The organic layers were washedwith sat. NH₄Cl, brine and dried over MgSO₄, filtered and concentrated.The crude product was used without purification for the next step.

[1406] Using same procedure for 554-RB-225, NY-21 (8 g, 95.1 mmol) wasconverted to NY-22 (27.33 g).

[1407] Using same procedure for NY-01, NY-20 (5.08 g, 19.2 mmol) wasconverted to NY-23 (4.22 g) as one of diastereomers.

[1408] Using same procedure for 343-yw-279, NY-23 (4.2 g, 7.23 mmol) wasconverted to NY-24 (3.7 g).

[1409] To a solution of NY-24 (3.69 g, 6.33 mmol) in CH₂Cl₂ (70 mL),2,6-lutidine (3.7 mL, 31.8 mmol) and TBSOTf (3.63 mL, 15.8 mmol) wereadded at 0° C. Then, the mixture was warmed to room temperature andstirred for 30 min. The mixtute was quenched with MeOH and poured intocold sat. NaHCO₃ and extracted with EtOAc. The organic layer was washedwith water, 5% citric acid, water, sat. NaHCO₃, brine and dried overNa₂SO₄, filtered and concentrated. The crude product was purified onsilica gel column with hexane/EtOAc, 100:1, 50:1, 30:1 to give 4.23 g ofNY-25

[1410] .

[1411] NY-25 (4.2 g, 6.03 mmol) was dissolved in dry Et₂O (50 mL) andthe solution was cooled to 0° C. in ice/water bath. Then LiBH₄ (135 mg,6.2 mmol) was added portionwise, the mixture was allowed to warm slowlyto rt and stirred for 2 days after which a saturated solution of NH₄Clwas added slowly. The mixture was extracted with EtOAc and the organicextract was washed with a saturated solution of NH₄Cl, water, brine,dried with anhydrous Na₂SO₄, filtered and concentrated. The crudeproduct was purified on silica gel column with hexane/EtOAc, 8:1 to give3.55 g of NY-26.

[1412] Using same procedure for 554-RB-260, NY-26 (568 mg, 0.927 mmol)was converted to NY-27 (565 mg).

[1413] Using same procedure for 10, NY-28 (2.7 g, 1 mmol) was convertedto NY-29 (2.45 g).

[1414] Using same procedure for 509-HD-212, NY-29 (2.45 g, 8.5 mmol) wasconverted to NY-11 (2.19 g).

[1415] Using same procedure for 509-HD-213, NY-30 (2.18 g, 3.06 mmol)was converted to NY-31 (2.35 g).

[1416] Using same procedure for 16, NY-27 (260 mg, 0.36 mmol) wasconverted to NY-32 (229 mg).

[1417] Using same procedure for 18, NY-32 (229 mg, 0.236 mmol) wasconverted to NY-33 (136 mg).

[1418] Using same procedure for 509-HD-116, NY-33 (136 mg, 0.585 mmol)was converted to NY-34(119 mg).

[1419] Using same procedure for TM-12, NY-34 (116 mg, 0.285 mmol) wasconverted to NY-35 (146 mg).

[1420] Using same procedure for TM-13, NY-35 (80 mg, 0.206 mmol) wasconverted to NY-36 (55 mg).

[1421] Using same procedure for NF-0675, NY-36 (52 mg, 0.135 mmol) wasconverted to NF-0934 (29 mg).

[1422] Synthetic procedure for NF-1418

[1423] Using same procedure for 10, NY-144 (7.11 g, 29.84 mmol) wasconverted to NY-145 (7.32 g).

[1424] Using same procedure for NY-45, NY-145 (7.32 g, 21.13 mmol) wasconverted to NY-146 (4.46 g).

[1425] Using same procedure for NY-48, NY-146 (318 mg, 1mmol) wasconverted to mixture of NY-147 and NY-148 (305 mg).

[1426] Using same procedure for 509-HD-213, NY-147 and NY-148 (303 mg,0.912 mmol) was converted to NY-149 (297 mg) and NY-150 (42 mg).

[1427] Using same procedure for 16, NY-53 (1.81 g, 4.808 mmol) wasconverted to NY-151 (2.06 g).

[1428] Using same procedure for 18, NY-151 (306 mg) was converted toNY-152 (250 mg).

[1429] Using same procedure for 509-HD-116, NY-152 (230 mg, 0.233 mmol)was converted to NY-153 (197 mg).

[1430] Using same procedure for TM-12, NY-153 (136 mg, 0.233 mmol) wasconverted to NY-154 (30 mg).

[1431] A mixture of NY-154 (28 mg, 0.0494 mmol), 1N NaOH (125 μL, 0.125mmol) and DME (0.5 mL) was stirred at room temperature for 18 hrs. Thereaction mixture was washed with Et₂O. The aqueous layer was neutralizedwith sat. NH₄Cl and extracted with EtOAc. The organic layers were washedwith brine, dried over Na₂SO₄, filtered and concentrated to give 23 mgof NY-155. NY-155 was used without purification for the next step.

[1432] A mixture of NY-155 (22 mg, 0.0398 mmol), diphenylphosphorylazide (8.6 μL, 0.0399 mmol), Et₃N (4 mg, 0.0395 mmol), ^(t)BuOH (0.3 mL)and toluene (1.5 mL) was refluxed for 3 hrs. The reaction mixture wasdiluted with EtOAc and washed with 5% citric acid, water, sat. NaHCO₃,brine, dried over Na₂SO₄, filtered and concentrated. The crude productwas purified on silica gel column with hexane/EtOAc, 5:1, 3:1 to give 12mg of NY-156.

[1433] Using same procedure for 509-HD-188, NY-156 (11 mg, 0.0176 mmol)was converted to NY-157 (7.5 mg).

[1434] Using same procedure for TM-13, NY-157 (7.5 mg, 0.0149 mmol) wasconverted to NY-158 (5.3 mg).

[1435] Using same procedure for NF-0675, NY-158 (5.2 mg, 0.0104 mmol)was converted to NF-1418 (4.7 mg).

[1436] Synthetic procedure for NF-1419

[1437] Using same procedure for B2538, NF-1418 (4 mg, 0.00937 mmol) wasconverted to NF-1419 (2.3 mg).

[1438] Measurement of Effect of Compounds on TNF-α and β-actin PLAP(Placental Alkaline Phosphatase) Transcription.

[1439] NF-κB is a critical nuclear factor to regulate various genesinvolved in immune and inflammatory responses. (see, Ghosh et al, AnnuRev Immunol. 1998, 16, 225). It is well characterized that TNFα genetranscription is regulated by NF-κB activation (see, Drouet et al. J.Immunol. 1991, 147, 1694), therefore, the assay with TNFα-PLAPtranscription was employed to evaluate the inhibitory effect of testcompounds on NF-κB activation.

[1440] A TNFα-PLAP plasmid (TNFα-promoter+5′-UTR (1.4 kb)+PLAP+SV40polyA+PGK-neo, Goto et al. Mol. Pharmacol. 1996, 49, 860) wasconstructed with slight modification in which TNFα-3′-UTR (772 b.p.) wasinserted between PLAP and SV40 polyA (TNFα-promoter+5′-UTR (1.4kb)+PLAP+TNFα-3′-UTR+SV40 polyA+PGKneo). Then the THP-1-33 cells wereestablished by stably transfecting the modified TNFcc-PLAP plasmid intoTHP-1 cells (human acute monocytic leukemia). In order to simultaneouslyevaluate non-specific effects of test compounds on transcription, B164cells were also established by stably transfecting β-actin-PLAP plasmid(β-actin-promoter (4.3 kb)+PLAP+SV40 polyA+PGKneo) into THP- 1 cells.THP-1-33 cells (TNFα-PLAP) produce PLAP activity by the stimulation withLPS; on the other hand, B164 cells (β-actin-PLAP) constantly producePLAP activity without stimuli.

[1441] THP-1-33 cells and B164 cells were maintained in RPMI1640containing 10% heat-inactivated endotoxin-free fetal bovine serum (FBS)and G418 (1 mg/ml). These cells were seeded at a density of 1.0×10⁴cells/well onto 96-well plate, and then were cultured in the presence orabsence of test compounds for 30 min, followed by stimulation with 100ng/mL of lipopolysaccharide (E. coli 0127:B08 or 01:B4). After thecultivation for 40-48 hrs, culture supernatant was harvested andalkaline phosphatase activity in the supernatant was measured.

[1442] Alkaline phosphatase activity was quantified with the use of achemiluminescent substrate,4-methoxy-4-(3-phosphatephenyl)spiro[1,2-dioxetane-3,2′-adamantane]. Toinactivate tissue-nonspecific alkaline phosphatase mainly derived fromFBS, samples were heated at 65° C. for 30 min before thechemiluminescent assay. Aliquots of 10 μL of culture supernatant weremixed with 50 μL of assay buffer (0.28 M Na₂CO₃—NaHCO₃, pH 10.0,containing 8 mM MgSO₄) in a 96-well Microlite™ plate (opaque), and then50 μL of chemiluminescent substrate was added and mixed. After 60 minincubation at room temperature, steady state chemiluminesce was measuredwith a microplate luminometer.

[1443] The PLAP activity of each sample was calculated as follows:

[1444] TNFα-PLAP % of control=(A−B)×100/(C−B)

[1445] β-actin-PLAP % of control=(A)×100/(C)

[1446] A: sample/chemiluminescence of the sample cultured with the testdrug & ed with LPS

[1447] B: blank/chemiluminescence of unstimulated sample

[1448] C: control/chemiluminescence of the sample cultured with LPS

[1449] The IC50 value of each test compound was calculated fromdose-inhibitory curve.

We claim:
 1. A pharmaceutical composition for systemic administrationcomprising a compound having the structure:

or pharmaceutically acceptable derivative thereof; wherein R₁ ishydrogen, aliphatic, heteroaliphatic, alicyclic, heteroalicyclic, arylor heteroaryl; R₂ and R₃ are each independently hydrogen, halogen,hydroxyl, protected hydroxyl, or an aliphatic, heteroaliphatic,alicyclic, heteroalicyclic, aryl or heteroaryl moiety; or R₁ and R₂,when taken together, may form a substituted or unsubstituted, saturatedor unsaturated cyclic ring of 3 to 8 carbon atoms; or R₁ and R₃, whentaken together, may form a substituted or unsubstituted, saturated orunsaturated cyclic ring of 3 to 8 carbon atoms; R4 is hydrogen orhalogen; R₅ is hydrogen, an oxygen protecting group or a prodrug; R₆ ishydrogen, hydroxyl, or protected hydroxyl; n is 0-2; R₇, for eachoccurrence, is independently hydrogen, hydroxyl, or protected hydroxyl;R₈ is hydrogen, halogen, hydroxyl, protected hydroxyl, alkyloxy, or analiphatic moiety optionally substituted with hydroxyl, protectedhydroxyl, SR₁₂, or NR₁₂R_(13;) R₉ is hydrogen, halogen, hydroxyl,protected hydroxyl, OR₁₂, SR₁₂, NR₁₂R₁₃, —X₁(CH₂)_(p)X₂—R₁₄, or is loweralkyl optionally substituted with hydroxyl, protected hydroxyl, halogen,amino, protected amino, or —X₁(CH₂)_(p)X₂—R₁₄; wherein R₁₂ and R₁₃ are,independently for each occurrence, hydrogen, aliphatic, heteroaliphatic,alicyclic, heteroalicyclic, aryl or heteroaryl; or a protecting group,or R₁₂ and R₁₃, taken together may form a saturated or unsaturatedcyclic ring containing 1 to 4 carbon atoms and 1 to 3 nitrogen or oxygenatoms, and each of R₁₂ and R₁₃ are optionally further substituted withone or more occurrences of hydroxyl, protected hydroxyl, alkyloxy,amino, protected amino, alkylamino, aminoalkyl, or halogen, wherein X₁and X₂ are each independently absent, or are oxygen, NH, or —N(alkyl),or wherein X₂—R₁₄ together are N₃ or are a saturated or unsaturatedheterocyclic moiety, p is 2-10, and R₁₄ is hydrogen, or an aryl,heteroaryl, alkylaryl, or alkylheteroaryl moiety, or is —(C═O)NHR₁₅—(C═O)OR₁₅, or —(C═O)R₁₅, wherein each occurrence of R₁₅ isindependently hydrogen, aliphatic, heteroaliphatic, alicyclic,heteroalicyclic, aryl or heteroaryl; or R₁₄ is —SO₂(R₁₆), wherein R₁₆ isan aliphatic moiety, wherein one or more of R₁₄, R₁₅, or R₁₆ areoptionally substituted with one or more occurrences of hydroxyl,protected hydroxyl, alkyloxy, amino, protected amino, alkylamino,aminoalkyl, or halogen; or R₈ and R₉ may, when taken together, form asaturated or unsaturated cyclic ring containing 1 to 4 carbon atoms and1 to 3 nitrogen or oxygen atoms and is optionally substituted withhydroxyl, protected hydroxyl, alkyloxy, amino, protected amino,alkylamino, aminoalkyl, or halogen; R₁₀ is hydrogen, hydroxyl, protectedhydroxyl, amino, or protected amino; R₁₁ is hydrogen, hydroxyl orprotected hydroxyl; X is absent or is O, NH, N-alkyl, CH₂ or S; Y isCHR₁₇, O, C═O, CR₁₇ or NR₁₇; and Z is CHR₁₈, O, C═O, CR₁₈ or NR₁₈,wherein each occurrence of R₁₇ and R₁₈ is independently hydrogen oraliphatic, or R₁₇ and R₁₈ taken together is —O—, —CH₂— or —NR₁₉—,wherein R₁₉ is hydrogen or lower alkyl, and Y and Z may be connected bya single or double bond; and a pharmaceutically suitable carrier ordiluent.
 2. The composition of claim 1, wherein: R₁ is hydrogen,straight or branched lower alkyl, straight or branched lowerheteroalkyl, or aryl, wherein the alkyl, heteroalkyl, and aryl groupsmay optionally be substituted with one or more occurrences of halogen,hydroxyl or protected hydroxyl; R₂ and R₃ are each independentlyhydrogen, halogen, hydroxyl, protected hydroxyl, straight or branchedlower alkyl, straight or branched lower heteroalkyl, or aryl, whereinthe alkyl, heteroalkyl, and aryl groups may optionally be substitutedwith one or more occurrences of halogen, hydroxyl or protected hydroxyl;or R₁ and R₂, when taken together, may form a saturated or unsaturatedcyclic ring of 3 to 8 carbon atoms, optionally substituted with one ormore occurrences of halogen; or R₁ and R₃, when taken together, may forma saturated or unsaturated cyclic ring of 3 to 8 carbon atoms,optionally substituted with one or more occurrences of halogen; R₄ ishydrogen or halogen; R₅ is hydrogen or a protecting group; R₆ ishydrogen, hydroxyl, or protected hydroxyl; n is 0-2; R₇, for eachoccurrence, is independently hydrogen, hydroxyl, or protected hydroxyl;R₈ is hydrogen, halogen, hydroxyl, protected hydroxyl, alkyloxy, orlower alkyl optionally substituted with hydroxyl, protected hydroxyl,SR₁₂, or NR₁₂R₁₃; R₉ is hydrogen, halogen, hydroxyl, protected hydroxyl,OR₁₂, SR₁₂, NR₁₂R₁₃, —X₁(CH₂)_(p)X₂—R₁₄, or is lower alkyl optionallysubstituted with hydroxyl, protected hydroxyl, halogen, amino, protectedamino, or —X₁(CH₂)_(p)X₂—R₁₄; wherein R₁₂ and R₁₃ are, independently foreach occurrence, hydrogen, lower alkyl, aryl, heteroaryl, alkylaryl, oralkylheteroaryl, or a protecting group, or R₁₂ and R₁₃, taken togethermay form a saturated or unsaturated cyclic ring containing 1 to 4 carbonatoms and 1 to 3 nitrogen or oxygen atoms, and each of R₁₂ and R₁₃ areoptionally further substituted with one or more occurrences of hydroxyl,protected hydroxyl, alkyloxy, amino, protected amino, alkylamino,aminoalkyl, or halogen, wherein X₁ and X₂ are each independently absent,or are oxygen, NH, or —N(alkyl), or wherein X₂—R₁₄ together are N₃ orare a saturated or unsaturated heterocyclic moiety, p is 2-10, and R₁₄is hydrogen, or an aryl, heteroaryl, alkylaryl, or alkylheteroarylmoiety, or is —(C═O)NHR₁₅ —C═O)OR₁₅, or —(C═O)R₁₅, wherein eachoccurrence of R₁₅ is independently hydrogen, alkyl, heteroalkyl, aryl,heteroaryl, alkylaryl, or alkylheteroaryl, or R₁₄ is —SO₂(R₁₆), whereinR₁₆ is an alkyl moiety, wherein one or more of R₁₄, R₁₅, or R₁₆ areoptionally substituted with one or more occurrences of hydroxyl,protected hydroxyl, alkyloxy, amino, protected amino, alkylamino,aminoalkyl, or halogen; or R₈ and R₉ may, when taken together, form asaturated or unsaturated cyclic ring containing 1 to 4 carbon atoms and1 to 3 nitrogen or oxygen atoms and is optionally substituted withhydroxyl, protected hydroxyl, alkyloxy, amino, protected amino,alkylamino, aminoalkyl, or halogen; R₁₀ is hydrogen, hydroxyl, protectedhydroxyl, amino, or protected amino; R₁₁ is hydrogen, hydroxyl orprotected hydroxyl; X is absent or is O, NH, N-alkyl, CH₂ or S; Y isCHR₁₇, O, C═O, CR₁₇ or NR₁₇; and Z is CHR₁₈, O, C═O, CR₁₈ or NR18,wherein each occurrence of R₁₇ and R₁₈ is independently hydrogen orlower alkyl, or R₁₇ and R₁₈ taken together is —O—, —CH₂— or —NR₁₉—,wherein R₁₉ is hydrogen or lower alkyl, and Y and Z may be connected bya single or double bond.
 3. The composition of claim 2, where X isoxygen and n is
 1. 4. The composition of claim 2, where R₄ is halogen.5. The composition of claim 2, where R₄ is fluorine.
 6. The compositionof claim 2, where Y and Z together represent-CH═CH—
 7. The compositionof claim 2, where Y and Z together represent trans —CH═CH—.
 8. Thecomposition of claim 2, wherein R₁ and R₂ are each methyl and R₃ ishydrogen and the compound has the structure:

wherein R₄-R₁₁, n, X, Y and Z are as defined in claim
 2. 9. Thecomposition of claim 8, wherein X is oxygen and n is
 1. 10. Thecomposition of claim 8, wherein R₄ is halogen.
 11. The composition ofclaim 8, wherein Y and Z together represent —CH═CH.
 12. The compositionof claim 8, wherein X is oxygen, n is 1, R₄ is halogen and Y and Ztogether represent —CH═CH—.
 13. The composition of claim 11 or 12wherein —CH═CH— is trans.
 14. The composition of claim 2, wherein R₉ isNR₁₂R₁₃ and the compound has the structure:

wherein R₁-R₁₂, n, X, Y and Z are as defined in claim 2, or R₁₃ and R₈may, when taken together, form a cyclic ring containing 1 to 4 carbonatoms and 1 to 3 nitrogen or oxygen atoms and is optionally substitutedwith hydrogen, alkyloxy, amino, alkylamino, aminoalkyl, and halogen. 15.The composition of claim 14, wherein X is oxygen and n is
 1. 16. Thecomposition of claim 14, wherein R₄ is halogen.
 17. The composition ofclaim 14, wherein Y and Z together represent —CH═CH—.
 18. Thecomposition of claim 14, wherein R₁ and R₂ are each methyl and R₃ ishydrogen.
 19. The composition of claim 14, wherein X is oxygen, n is 1,R₁ and R₂ are each methyl, R₃ is hydrogen, R₄ is halogen, and Y and Ztogether represent —CH═CH—.
 20. The composition of claim 17 or 19,wherein —CH═CH— is trans.
 21. The composition of claim 1 wherein thecompound has the structure:

or pharmaceutically acceptable derivative thereof.
 22. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 23. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 24. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 25. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 26. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 27. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 28. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 29. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 30. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 31. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 32. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 33. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 34. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 35. The compositionof claim 1 wherein the compound has the structure:

or pharmaceutically acceptable derivative thereof.
 36. Thepharmaceutical composition of claim 1, wherein the composition is fororal administration.
 37. The pharmaceutical composition of claim 1,wherein the compound is present in an amount effective to inhibitproduction of a pro-inflammatory and/or immunologic cytokine.
 38. Thepharmnaceutical composition of claim 37, wherein the pro-inflammatoryand/or immunologic cytokine is TNFα, IL-1, IL-6, IL-8 or IL-2.
 39. Amethod for treating an inflammatory and/or autoimmune disordercomprising: systemically administering to a subject in need thereof atherapeutically effective amount of a compound having the structure:

or pharmaceutically acceptable derivative thereof; wherein R₁ ishydrogen, aliphatic, heteroaliphatic, alicyclic, heteroalicyclic, arylor heteroaryl; R₂ and R₃ are each independently hydrogen, halogen,hydroxyl, protected hydroxyl, or an aliphatic, heteroaliphatic,alicyclic, heteroalicyclic, aryl or heteroaryl moiety; or R₁ and R₂,when taken together, may form a substituted or unsubstituted, saturatedor unsaturated cyclic ring of 3 to 8 carbon atoms; or R₁ and R₃, whentaken together, may form a substituted or unsubstituted, saturated orunsaturated cyclic ring of 3 to 8 carbon atoms; R₄ is hydrogen orhalogen; R₅ is hydrogen, an oxygen protecting group or a prodrug; R₆ ishydrogen, hydroxyl, or protected hydroxyl; n is 0-2; R₇, for eachoccurrence, is independently hydrogen, hydroxyl, or protected hydroxyl;R₈ is hydrogen, halogen, hydroxyl, protected hydroxyl, alkyloxy, or analiphatic moiety optionally substituted with hydroxyl, protectedhydroxyl, SR₁₂, or NR₁₂R₁₃; R₉ is hydrogen, halogen, hydroxyl, protectedhydroxyl, OR₁₂, SR₁₂, NR₁₂R₁₃, —X₁(CH₂)_(p)X₂—R₁₄, or is lower alkyloptionally substituted with hydroxyl, protected hydroxyl, halogen,amino, protected amino, or —X₁(CH₂)_(p)X₂—R₁₄; wherein R₁₂ and R₁₃ are,independently for each occurrence, hydrogen, aliphatic, heteroaliphatic,alicyclic, heteroalicyclic, aryl or heteroaryl; or a protecting group,or R₁₂ and R₁₃, taken together may form a saturated or unsaturatedcyclic ring containing 1 to 4 carbon atoms and 1 to 3 nitrogen or oxygenatoms, and each of R₁₂ and R₁₃ are optionally further substituted withone or more occurrences of hydroxyl, protected hydroxyl, alkyloxy,amino, protected amino, alkylamino, aminoalkyl, or halogen, wherein X₁and X₂ are each independently absent, or are oxygen, NH, or —N(alkyl),or wherein X₂—R₁₄ together are N₃ or are a saturated or unsaturatedheterocyclic moiety, p is 2-10, and R₁₄ is hydrogen, or an aryl,heteroaryl, alkylaryl, or alkylheteroaryl moiety, or is —(C═O)NHR₁₅—(C═O)OR₁₅, or —(C═O)R₁₅, wherein each occurrence of R₁₅ isindependently hydrogen, aliphatic, heteroaliphatic, alicyclic,heteroalicyclic, aryl or heteroaryl; or R₁₄ is —SO₂(R₁₆), wherein R₁₆ isan aliphatic moiety, wherein one or more of R₁₄, R₁₅ or R₁₆ areoptionally substituted with one or more occurrences of hydroxyl,protected hydroxyl, alkyloxy, amino, protected amino, alkylamino,aminoalkyl, or halogen; or R₈ and R₉ may, when taken together, form asaturated or unsaturated cyclic ring containing 1 to 4 carbon atoms and1 to 3 nitrogen or oxygen atoms and is optionally substituted withhydroxyl, protected hydroxyl, alkyloxy, amino, protected amino,alkylamino, aminoalkyl, or halogen; R₁₀ is hydrogen, hydroxyl, protectedhydroxyl, amino, or protected amino; R₁₁ is hydrogen, hydroxyl orprotected hydroxyl; X is absent or is O, NH, N-alkyl, CH₂ or S; Y isCHR₁₇, O, C═O, CR₁₇ or NR₁₇; and Z is CHR₁₈, O, C═O, CR₁₈ or NR₁₈,wherein each occurrence of R₁₇ and R₁₈ is independently hydrogen oraliphatic, or R₁₇ and R₁₈ taken together is —O—, —CH₂— or —NR₁₉—,wherein R₁₉ is hydrogen or lower alkyl, and Y and Z may be connected bya single or double bond; and a pharmaceutically acceptable carrier ordiluent.
 40. The method of claim 39, wherein the compound isadministered orally.
 41. The method of claim 39 or 40, wherein themethod is for treating a disorder selected from the group consisting ofrheumatoid arthritis, psoriasis, asthma, cancer, sepsis, inflammatorybowel disease, atopic dermatitis, Crohn's disease, and autoimmunedisorders.
 42. The method of claim 41, wherein the method is fortreating psoriasis.
 43. The method of claim 41, wherein thecompound hasthe structure:

or pharmaceutically acceptable derivative thereof.
 44. The method ofclaim 39, wherein the compound is present in an amount effective toinhibit production of a pro-inflammatory and/or immunologic cytokine.45. The method of claim 44, wherein the pro-inflammatory and/orimmunologic cytokine is TNFα, IL-1, IL-6, IL-8 or IL-2.